Beyond thermoregulation: metabolic function of cetacean blubber in migrating bowhead and beluga whales.
*Lipid Metabolism; Adipose Tissue/*metabolism; Aging/metabolism; Amino Acid Sequence; Animals; Base Sequence; Beluga Whale/*physiology; Blubber; Body Temperature Regulation; Bowhead whale; Bowhead Whale/*physiology; Development; Female; Humans; Inbred C57BL; Leptin; Leptin/genetics; Leptin/genetics/metabolism; Lipase/genetics; Long-Evans; Male; Metabolic activity; Mice; Rats; Receptors; Seasons
The processes of lipid deposition and utilization, via the gene leptin (Lep), are poorly understood in taxa with varying degrees of adipose storage. This study examines how these systems may have adapted in marine aquatic environments inhabited by cetaceans. Bowhead (Balaena mysticetus) and beluga whales (Delphinapterus leucas) are ideal study animals-they possess large subcutaneous adipose stores (blubber) and undergo bi-annual migrations concurrent with variations in food availability. To answer long-standing questions regarding how (or if) energy and lipid utilization adapted to aquatic stressors, we quantified variations in gene transcripts critical to lipid metabolism related to season, age, and blubber depth. We predicted leptin tertiary structure conservation and assessed inter-specific variations in Lep transcript numbers between bowheads and other mammals. Our study is the first to identify seasonal and age-related variations in Lep and lipolysis in these cetaceans. While Lep transcripts and protein oscillate with season in adult bowheads reminiscent of hibernating mammals, transcript levels reach up to 10 times higher in bowheads than any other mammal. Data from immature bowheads are consistent with the hypothesis that short baleen inhibits efficient feeding. Lipolysis transcripts also indicate young Fall bowheads and those sampled during Spring months limit energy utilization. These novel data from rarely examined species expand the existing knowledge and offer unique insight into how the regulation of Lep and lipolysis has adapted to permit seasonal deposition and maintain vital blubber stores.
Ball H C; Londraville R L; Prokop J W; George John C; Suydam R S; Vinyard C; Thewissen J G M; Duff R J
Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology
2017
2017-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s00360-016-1029-6" target="_blank" rel="noreferrer noopener">10.1007/s00360-016-1029-6</a>
Intrastrain variants of herpes simplex virus type 1 isolated from a neonate with fatal disseminated infection differ in the ICP34.5 gene, glycoprotein processing, and neuroinvasiveness.
Humans; Male; Animals; Mice; Amino Acid Sequence; *Genetic Variation; Base Sequence; Molecular Sequence Data; Polymerase Chain Reaction/methods; DNA; Deoxyribonuclease BamHI; Deoxyribonuclease EcoRI; Glycoproteins/metabolism; Viral Envelope Proteins/analysis; Viral Proteins/*genetics; Genes; Viral; Animal; Disease Models; Herpesvirus 1; Inbred BALB C; Amino Acid; Sequence Homology; Sequence Analysis; Nucleic Acid; Polymorphism; *Protein Processing; Post-Translational; Human/*genetics/growth & development/isolation & purification; Restriction Fragment Length
Two intrastrain variants of herpes simplex virus type 1 (HSV-1) were isolated from a newborn with fatal disseminated infection. A small-plaque-producing variant (SP7) was the predominant virus (\textgreater99%) in the brain, and a large-plaque-producing variant (LP5) was the predominant virus (\textgreater99%) in the lung and gastrointestinal tract. EcoRI and BamHI restriction fragment patterns indicated that SP7 and LP5 are related strains. The large-plaque variants produced plaques similar in size to those produced by HSV-1 KOS. Unlike LP5 or KOS, SP7 was highly cell associated and processing of glycoprotein C and glycoprotein D was limited to precursor forms in infected Vero cells. The large-plaque phenotype from KOS could be transferred into SP7 by cotransfection of plasmids containing the EK or JK EcoRI fragment or a 3-kb plasmid with the UL34.5 gene of HSV-1 KOS together with SP7 DNA. PCR analysis using primers from within the ICP34.5 gene indicated differences for SP7, LP5, and KOS. Sequencing data indicated two sets of deletions in the UL34.5 gene that distinguish SP7 from LP5. Both SP7 and LP5 variants were neurovirulent (lethal following intracranial inoculation of young BALB/c mice); however, the LP5 variant was much less able to cause lethal neuroinvasive disease (footpad inoculation) whereas KOS caused no disease. Passage of SP7 selected for viruses (SLP-5 and SLP-10) which were attenuated for lethal neuroinvasive disease, were not cell-associated, and differed in the UL34.5 gene. UL34.5 from SLP-5 or SLP-10 resembled that of KOS. These findings support a role for UL34.5 in promoting virus egress and for neuroinvasive disease.
Bower J R; Mao H; Durishin C; Rozenbom E; Detwiler M; Rempinski D; Karban T L; Rosenthal K S
Journal of virology
1999
1999-05
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
Calcium channel blockade attenuates angiotensin II-induced drinking in rats.
Amino Acid Sequence; Angiotensin II/*antagonists & inhibitors/pharmacology; Animals; Calcium Channel Blockers/administration & dosage/*pharmacology; Dimethyl Sulfoxide/pharmacology; Dose-Response Relationship; Drinking Behavior/*drug effects; Drug; Injections; Intraventricular; Isradipine/pharmacology; Male; Molecular Sequence Data; Rats; Sprague-Dawley
Lateral ventricular administration of angiotensin II (ANG II) produces potent dipsogenic effects in water-sated rats. ANG II seems to require functional voltage-gated calcium channels on neurons throughout circumventricular brain sites to exert its effects. Although there are at least three types of calcium channels, only L-type calcium channel-blocking drugs have been reported to decrease drinking. (4-(4-Benzofurazanyl)-1-4-dihydro-2,6-dimethyl-3,5-pyridine-dic arb oxylic acid methyl 1-methyl-ethyl ester) [PN 200-110; isradipine (ISR)], a selective L-type calcium channel blocker, has been shown to attenuate significantly the intake of sweetened water in water-sated rats following either peripheral or ICV administration, but ISR does not affect plain-water intake in water-deprived rats. The present experiment was designed to determine whether ISR would attenuate ANG II-induced drinking that is not either motivated by palatability or dependent on deprivation. Rats, each fitted with chronic indwelling ventricular cannulae, were pretreated with ISR (0.3, 3.0, and 30 micrograms/rat; ICV). ANG II (40 ng/rat; ICV) was administered 10 min later and rats were allowed free access to water for 15 min. Injections of ANG II plus saline and ANG II plus the ISR vehicle (dimethyl sulfoxide) did not attenuate ANG II-induced polydipsia, whereas ANG II+ISR (0.3 and 3.0 micrograms) attenuated ANG II-induced drinking to 62 and 22% of control, respectively. Results with the 30-micrograms dose were not different from the 3.0 dose.(ABSTRACT TRUNCATED AT 250 WORDS)
Calcagnetti D J; Schechter M D
Brain research bulletin
1993
1905-6
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/0361-9230(93)90087-r" target="_blank" rel="noreferrer noopener">10.1016/0361-9230(93)90087-r</a>
Identification of a new P450 subfamily, CYP4F1, expressed in rat hepatic tumors.
2-Acetylaminofluorene; Aflatoxin B1; Amino Acid; Amino Acid Sequence; Animals; Antisense Elements (Genetics); Base Sequence; Blotting; Clofibrate; Cytochrome P-450 Enzyme System/analysis/*genetics; Humans; Liver Neoplasms/chemically induced/*enzymology/genetics; Male; Microsomes/enzymology; Molecular Sequence Data; Multigene Family; Northern; Rats; Sequence Homology; Western
The expression of the rat cytochrome P450 CYP4 family was studied in hepatic tumors. In most of the primary and transplantable hepatic tumors studied, lauric acid omega-hydroxylase activity associated with the CYP4A subfamily enzymes decreased. The expression of CYP4A proteins and mRNAs in these tumors as assessed by Western and Northern blot was undetectable. However, while RNA analysis revealed the absence of 4A1, 4A2, and 4A3 mRNAs, the expression of CYP4 gene(s) was detected. A Uni-ZAP cDNA library was constructed from a
Chen L; Hardwick J P
Archives of biochemistry and biophysics
1993
1993-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1006/abbi.1993.1003" target="_blank" rel="noreferrer noopener">10.1006/abbi.1993.1003</a>
Cloning and 5'-flanking sequence of a rat cholesterol 7 alpha-hydroxylase gene.
Animals; Rats; Amino Acid Sequence; Base Sequence; Molecular Sequence Data; Cholesterol 7-alpha-Hydroxylase/*genetics; DNA; Restriction Mapping; Genetic; Cloning; Molecular; *Promoter Regions
A cholesterol 7 alpha-hydroxylase gene containing 8 kb of the 5'-flanking region and 5 kb of the transcription region which covers exons 1 to 5 was isolated from a rat genomic library. The 2015 bp nucleotide sequence 5'-upstream from the start codon was determined. This promotor region contains many liver-enriched or -specific elements (TGT3, HNF/LF-B1), putative hormone responsive elements (TRE, GRE, RRE or RARE) and ubiquitous transcription factor binding sites (NF-1,
Chiang J Y; Yang T P; Wang D P
Biochimica et biophysica acta
1992
1992-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
Genomic cloning, sequencing, and analysis of the hamster cholesterol 7 alpha-hydroxylase gene (CYP7).
Amino Acid; Amino Acid Sequence; Animals; Base Sequence; Blotting; Cholesterol 7-alpha-Hydroxylase/*genetics; Cloning; Cricetinae; DNA/genetics/isolation & purification; Genomic Library; Humans; Liver/enzymology; Mesocricetus/*genetics; Messenger/biosynthesis/metabolism; Molecular; Molecular Sequence Data; Northern; Poly A/biosynthesis/metabolism; Rats; Restriction Mapping; RNA; Sequence Homology; Southern
Cholesterol 7 alpha-hydroxylase is the rate limiting enzyme in bile acid biosynthesis and plays an important role in cholesterol homeostasis. The Golden Syrian hamster has been used as an animal model for the study of atherosclerosis and cholesterol gallstone disease. We have screened a lambda DASH II hamster liver genomic library using a rat cDNA as a hybridization probe. A 14-kb genomic clone has been isolated and characterized by restriction mapping and Southern blot hybridization. The clone contained the full-length gene encoding cholesterol 7 alpha-hydroxylase together with an upstream sequence of approximately 5 kb. DNA sequencing and analysis of about 11 kb of the gene revealed that the hamster CYP7 gene consists of six exons and five introns, which have the same structures and sizes as predicted in the rat and human CYP7 genes. The nucleotide and deduced amino acid sequences of the hamster cholesterol 7 alpha-hydroxylase have a high sequence identity of about 90% to the rat and 82% to the human sequences. Particularly, exons 2, 5, and 6 are highly conserved among these species, thus reflecting the presence of some domains that are crucial for the activity of this unique enzyme. The putative cholesterol-binding region, an aromatic amino acid region, and the P450 heme-binding region are completely conserved. Comparison of the 250-bp 5'-flanking sequence to the corresponding region in the rat and human genes revealed a high degree of homology ranging between 71% and 82%. Next to the canonical TATA and CCAAT boxes are many consensus sequences (LF-A1, LF-B1, TGT3) for liver-specific or -enriched transcription factors (HNF4, HNF1, and HNF5, respectively) and an imperfect direct repeat of thyroid hormone responsive element (TRE), which is located between TGT3 and LF-B1. These sequence motifs are completely conserved among the rat, human, and hamster CYP7 genes. Several modified sterol regulatory element (SRE)-like sequences are located in the upstream flanking region and in the first intron. This highly conserved proximal promoter may play important roles in the transcription activity and in the regulation of the CYP7 gene by physiological agents, such as bile acids and steroid/thyroid hormones. This is the first report describing the complete nucleotide sequence and confirming the structure of a CYP7 gene.(ABSTRACT TRUNCATED AT 400 WORDS)
Crestani M; Galli G; Chiang J Y
Archives of biochemistry and biophysics
1993
1993-11
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1006/abbi.1993.1537" target="_blank" rel="noreferrer noopener">10.1006/abbi.1993.1537</a>
The isolation and characterization of the bovine cytochrome b5 gene, and a transcribed pseudogene.
*Pseudogenes; Amino Acid Sequence; Animals; Base Sequence; Blotting; Cattle/*genetics; Cytochromes b5/*genetics; DNA/genetics/isolation & purification; Exons; Gene Expression; Genetic; Genomic Library; Humans; Introns; Messenger/biosynthesis/metabolism; Molecular Sequence Data; Nucleic Acid; Nucleic Acid Conformation; Oligodeoxyribonucleotides; Rabbits; Restriction Mapping; Reticulocytes/metabolism; RNA; RNA Precursors/chemistry/metabolism; Sequence Homology; Southern; Transcription
This is the first isolation and characterization of a cytochrome b5(b5) gene. The bovine b5 gene is quite large, spanning about 28 kb and contains six exons. One of these exons appears to code for a reticulocyte-specific sequence similar to that described for human and rabbit b5. All of the splicing junctions conform to the GT-AG consensus rule. The 5' flanking sequence has no obvious TATA box, two CAAT boxes, and contains several G:C-rich SpI motifs indicative of a house-keeping gene. In reticulocyte mRNA we found evidence for a transcribed b5 pseudogene, but could not detect sequences coding for the soluble form of b5. We conclude that the soluble form of b5 is derived from the membrane-bound b5 by a post-translational mechanism.
Cristiano R J; Giordano S J; Steggles A W
Genomics
1993
1993-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1006/geno.1993.1331" target="_blank" rel="noreferrer noopener">10.1006/geno.1993.1331</a>
The complete nucleotide sequence of bovine liver cytochrome b5 mRNA.
Animals; Amino Acid Sequence; Base Sequence; Molecular Sequence Data; Liver/*enzymology; Cattle; Cytochromes b5; Cytochrome b Group/*genetics/isolation & purification; RNA; Messenger/*isolation & purification
Cristiano R J; Steggles A W
Nucleic acids research
1989
1989-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1093/nar/17.2.799" target="_blank" rel="noreferrer noopener">10.1093/nar/17.2.799</a>
Isolation and characterization of cDNA clones for human, bovine and rabbit liver cytochrome b5 mRNA's.
Humans; Animals; Amino Acid Sequence; Rabbits; Base Sequence; Molecular Sequence Data; Liver/*enzymology; Cattle; Chickens; Cytochromes b5/blood/*genetics; DNA/*isolation & purification; Erythrocytes/enzymology; RNA; Sequence Homology; Cloning; Molecular; Nucleic Acid; Messenger/*genetics
Cristiano R J; Yoo M; Dariush N; Steggles A W
Progress in clinical and biological research
1989
1905-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
The nucleotide sequence of rabbit liver cytochrome b5 mRNA.
Animals; Amino Acid Sequence; Liver/*metabolism; Rabbits; Base Sequence; Molecular Sequence Data; Cytochrome b Group/*genetics; Cytochromes b5; Chromosome Deletion; Genes; RNA; Cloning; Molecular; Messenger/*genetics
Using a synthetic 26 base pair (bp) oligonucleotide, we have screened a rabbit liver cDNA library in lambda gt11 and isolated a 1000 bp DNA clone corresponding to cytochrome b5 mRNA. The clone contains the complete coding region and long 5' and 3' non-translated regions. The derived amino acid sequence (sequence; see text) agrees with published sequences, and confirms that the amino acids 62 and 104 are asparagine and aspartic acid, respectively.
Dariush N; Fisher C W; Steggles A W
Protein sequences & data analysis
1988
1905-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
The human liver and reticulocyte cytochrome b5 mRNAs are products from a single gene.
*Genes; Amino Acid Sequence; Base Sequence; Cytochromes b5/blood/*genetics; Gene Library; Genetic; Humans; Liver/*metabolism; Molecular Sequence Data; Polymerase Chain Reaction; Polymorphism; Reticulocytes/*metabolism; RNA/blood/genetics/isolation & purification
Using a combination of standard cDNA library screening techniques and the Polymerase Chain Reaction (PCR) we have isolated and sequenced DNA fragments corresponding to the human reticulocyte cytochrome b5 mRNA. The reticulocyte specific sequence codes for amino acids 97 and 98 only, with a TAA stop codon. The reticulocyte specific 3'non-translated sequence has 15 new nucleotides then utilizes the liver mRNA sequence from amino acid 97 onward. This indicates that the reticulocyte specific exon has 24 base pairs (bp). In addition, we have isolated sequences that are derived from a transcribed cytochrome b5 pseudogene. This transcript contains multiple mutations which prevent the synthesis of any functional protein.
Giordano S J; Steggles A W
Biochemical and biophysical research communications
1991
1991-07
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<a href="http://doi.org/10.1016/0006-291x(91)91776-9" target="_blank" rel="noreferrer noopener">10.1016/0006-291x(91)91776-9</a>
Differential expression of the mRNAs for the soluble and membrane-bound forms of rabbit cytochrome b5.
Agar Gel; Amino Acid Sequence; Animals; Base Sequence; Blotting; Cell Membrane/metabolism; Cytochromes b5/*genetics; Cytosol/metabolism; DNA/genetics/isolation & purification; Electrophoresis; Exons; Leukocytes/metabolism; Liver/*metabolism; Messenger/genetics/isolation & purification/*metabolism; Molecular Sequence Data; Oligodeoxyribonucleotides; Polymerase Chain Reaction/methods; Rabbits; Reticulocytes/metabolism; RNA; Southern
Total RNA was extracted from a variety of rabbit tissues and reverse transcribed for use in the polymerase chain reaction technique. Using primers designed to amplify the membrane-bound liver cytochrome b5 cDNA, products of two sizes were observed. Both hybridized strongly to a radiolabelled liver cytochrome b5 probe. Sequencing confirmed that the two types of cDNA product encoded the membrane-bound and the soluble forms of b5. Messenger RNA corresponding to the soluble cytochrome was detected in the lung, gallbladder and the adrenal gland, as well as in reticulocytes and bone marrow. This was an unexpected finding since the protein has been isolated only from erythrocytes. In contrast, membrane-bound cytochrome b5 mRNA was detected in all tissues tested, suggesting that the corresponding protein is ubiquitous in tissue distribution.
Giordano S J; Steggles A W
Biochimica et biophysica acta
1993
1993-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/0167-4781(93)90274-h" target="_blank" rel="noreferrer noopener">10.1016/0167-4781(93)90274-h</a>
Identification and nucleotide sequence of the leukocyte and reticulocyte forms of rabbit cytochrome b5 mRNA.
Animals; Amino Acid Sequence; Rabbits; Base Sequence; Polymerase Chain Reaction; Molecular Sequence Data; Nucleic Acid Hybridization; Cytochromes b5/chemistry/*genetics; Leukocytes/*chemistry; Liver/chemistry; Reticulocytes/*chemistry; RNA; Sequence Homology; Nucleic Acid; Messenger/*chemistry
RNA extracted from rabbit leukocytes and reticulocytes was reverse transcribed and used in the Polymerase Chain Reaction technique along with primers designed to amplify the coding sequence of rabbit cytochrome b5. The resultant amplified products were subcloned and analyzed. Sequencing confirmed that leukocyte and liver cDNAs are homologous and encode the membrane-bound form of the protein. In contrast, reticulocytes exhibit a highly similar, but different mRNA which encodes the smaller, soluble cytochrome b5. This is the first example of a cytochrome b5 sequence from a tissue other than liver, erythrocyte or reticulocyte.
Giordano S J; Steggles A W
SAAS bulletin, biochemistry and biotechnology
1992
1992-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
A L.E.A.P.S. heteroconjugate vaccine containing a T cell epitope from HSV-1 glycoprotein D elicits Th1 responses and protection.
Amino Acid Sequence; Animals; CD4-Positive T-Lymphocytes/immunology; CD8-Positive T-Lymphocytes/immunology; Conjugate/immunology; Delayed/immunology; Enzyme-Linked Immunosorbent Assay; Epitopes/*immunology; Female; Herpes Simplex Virus Vaccines/*immunology; Herpes Simplex/pathology/prevention & control; Herpesvirus 1; Human/*immunology; Hypersensitivity; Inbred BALB C; Interferon-gamma/biosynthesis; Lymphocyte Count; Mice; Molecular Sequence Data; Peptides/immunology; Th1 Cells/*immunology; Vaccines; Viral Envelope Proteins/*immunology
The L.E.A.P.S. heteroconjugate vaccine antigen (JgD), composed of a T cell epitope from glycoprotein D (gD(8-23)) of herpes simplex virus (HSV) linked with a peptide sequence from beta-2-microglobulin (aa38-50), elicited protection against lethal intraperitoneal (IP) challenge and prevented disease signs in most, and limited disease progression, for the rest of BALB/c mice challenged in the epidermal abrasion-zosteriform spread mouse infection model. JgD elicited a Th1 response in vaccinated mice as indicated by delayed type hypersensitivity (DTH) responses to HSV antigen, and gD and virion specific antibodies with an IgG2a/IgG1 \textgreater1. Vaccination with the JgD peptide delayed the onset of disease signs, reduced severity of the disease and reduced mortality rates in mice with different MHC backgrounds as compared to their respective control mice. CD8 cells were demonstrated as important for initiation of the immune response to JgD and CD4 cells and interferon gamma (IFN-gamma) for delivering immune protection in BALB/c mice, as indicated in monoclonal antibody ablation studies. JgD, and other
Goel N; Rong Q; Zimmerman D; Rosenthal K S
Vaccine
2003
2003-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/s0264-410x(03)00429-8" target="_blank" rel="noreferrer noopener">10.1016/s0264-410x(03)00429-8</a>
Increased serotransferrin and ceruloplasmin turnover in diet-controlled patients with type 2 diabetes.
*Ceruloplasmin; *Deamidation; *Heavy water metabolic labeling; *High resolution mass spectrometry; *Iron metabolism; *LC-MS/MS; *Non-enzymatic glycation; *Oxidative stress; *Protein Processing; *Proteome dynamics; *Serotransferrin; *Type 2 diabetes mellitus; Adult; Amino Acid Sequence; Case-Control Studies; Ceruloplasmin/genetics/*metabolism; Deuterium/metabolism; Diabetes Mellitus; Diabetic; Diet; Female; Gene Expression Regulation; Glycated Hemoglobin A/genetics/metabolism; Glycosylation; Humans; Iron/*metabolism; Isotope Labeling; Male; Middle Aged; Oxidation-Reduction; Oxidative Stress; Post-Translational; Proteolysis; Transferrin/genetics/*metabolism; Type 2/diet therapy/genetics/*metabolism/pathology
Type 2 diabetes mellitus (T2DM) is associated with oxidative stress and perturbed iron metabolism. Serotransferrin (Trf) and ceruloplasmin (Cp) are two key proteins involved in iron metabolism and anti-oxidant defense. Non-enzymatic glycation and oxidative modification of plasma proteins are known to occur under hyperglycemia and oxidative stress. In this study, shotgun proteomics and (2)H2O-based metabolic labeling were used to characterize post-translational modifications and assess the kinetics of Trf and Cp in T2DM patients and matched controls in vivo. Six early lysine (Amadori) and one advanced arginine glycation were detected in Trf. No glycation, but five asparagine deamidations, were found in Cp. T2DM patients had increased fractional catabolic rates of both Trf and Cp that correlated with HbA1c (p \textless 0.05). The glycated Trf population was subject to an even faster degradation compared to the total Trf pool, suggesting that hyperglycemia contributed to an increased Trf degradation in T2DM patients. Enhanced production of Trf and Cp kept their levels stable. The changes in Trf and Cp turnover were associated with increased systemic oxidative stress without any alteration in iron status in T2DM. These findings can help better understand the potential role of altered Trf and Cp metabolism in the pathogenesis of T2DM and other diseases.
Golizeh Makan; Lee Kwangwon; Ilchenko Serguei; Osme Abdullah; Bena James; Sadygov Rovshan G; Kashyap Sangeeta R; Kasumov Takhar
Free radical biology & medicine
2017
2017-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.freeradbiomed.2017.10.373" target="_blank" rel="noreferrer noopener">10.1016/j.freeradbiomed.2017.10.373</a>
Specificity of metabotropic glutamate receptor 2 coupling to G proteins.
Amino Acid; Amino Acid Sequence; Animals; Calcium/metabolism; Electrophysiology; Gi-Go/metabolism; GTP-Binding Protein alpha Subunits; GTP-Binding Proteins/*metabolism; Metabotropic Glutamate/*metabolism; Molecular Sequence Data; Neurons/*drug effects/metabolism; Pertussis Toxin/*pharmacology; Rats; Receptors; Sequence Homology; Superior Cervical Ganglion/cytology; Wistar
Metabotropic glutamate receptor 2 (mGluR2) is a class 3 G protein-coupled receptor and an important mediator of synaptic activity in the central nervous system. Previous work demonstrated that mGluR2 couples to pertussis toxin (PTX)-sensitive G proteins. However, the specificity of mGluR2 coupling to individual members of the G(i/o) family is not known. Using heterologously expressed mGluR2 in rat sympathetic neurons from the superior cervical ganglion (SCG), the mGluR2/G protein coupling profile was characterized by reconstituting coupling in PTX-treated cells expressing PTX-insensitive mutant Galpha proteins and Gbetagamma. By employing this method, it was demonstrated that mGluR2 coupled strongly with Galphaob, Galphai1, Galphai2, and Galphai3, although coupling to Galphaoa was less efficient. In addition, mGluR2 did not seem to couple to the most divergent member of the G(i/o) family, Galphaz, although Galphaz coupled strongly to the endogenous alpha2 adrenergic receptor. To determine which Galpha proteins may be natively expressed in SCG neurons, the presence of mRNA for various Galpha proteins was tested using reverse transcription-polymerase chain reaction. Strong bands were detected for all members of the G(i/o) family (Galphao, Galphai1, Galphai2, Galphai3, Galphaz) as well as for Galpha11 and Galphas. A weak signal was detected for Galphaq and no Galpha15 mRNA was detected.
Kammermeier Paul J; Davis Margaret I; Ikeda Stephen R
Molecular pharmacology
2003
2003-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1124/mol.63.1.183" target="_blank" rel="noreferrer noopener">10.1124/mol.63.1.183</a>
Protein thiyl radical mediates S-glutathionylation of complex I.
*Oxidative Stress; Amino Acid Motifs; Amino Acid Sequence; Animals; Binding Sites; Cattle; Cell Line; Cyclic N-Oxides/chemistry/pharmacology; Cysteine/chemistry/*metabolism; Electron Transport Complex I/chemistry/*metabolism; Free Radical Scavengers/chemistry/pharmacology; Free Radicals/chemistry/*metabolism; Glutathione/chemistry/*metabolism; Heart/enzymology/metabolism; Mice; Mitochondria; Models; Molecular; Molecular Sequence Data; Muscle Cells/drug effects/metabolism; Onium Compounds/pharmacology; Peptide Fragments/chemistry; Peptide Mapping; Protein; Rats; Rotenone/pharmacology; Structural Homology; Superoxides/metabolism
Complex I is a critical site of O(2)(*-) production and the major host of reactive protein thiols in mitochondria. In response to oxidative stress, complex I protein thiols at the 51- and 75-kDa subunits are reversibly
Kang Patrick T; Zhang Liwen; Chen Chwen-Lih; Chen Jingfeng; Green Kari B; Chen Yeong-Renn
Free radical biology & medicine
2012
2012-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.freeradbiomed.2012.05.025" target="_blank" rel="noreferrer noopener">10.1016/j.freeradbiomed.2012.05.025</a>
Polymorphisms of human cholesterol 7 alpha-hydroxylase.
Humans; Amino Acid Sequence; Base Sequence; Polymerase Chain Reaction; Molecular Sequence Data; Cholesterol 7-alpha-Hydroxylase/*genetics; DNA/genetics; Restriction Mapping; Oligodeoxyribonucleotides/chemistry; Genetic; Cloning; Molecular; Polymorphism
Human liver cholesterol 7 alpha-hydroxylase (CYP7) cDNAs were isolated from a human liver cDNA library. A full-length cDNA has 2901 nucleotides which encode a typical P450 polypeptide of 504 amino acid residues. Two different sequences of codon 100, TTT (Phe) and TCT (Ser), were identified in cDNA clones. In addition, codons 347 and 385 are GAT (Asp) and GAC (Asp) in all cDNA clones, whereas those reported previously (FEBS Lett. 268, 137-140, 1990) are AAT (Asn) and AGC (Ser), respectively. Since there is only one 7 alpha-hydroxylase gene in the human genome, it is likely that polymorphisms at the codon 100 of cDNA clones arise from two different alleles in the 7 alpha-hydroxylase gene of this human liver.
Karam W G; Chiang J Y
Biochemical and biophysical research communications
1992
1992-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
Generation and characterization of ATP analog-specific protein kinase Cdelta.
Adenosine Triphosphate/*analogs & derivatives/*chemistry; Amino Acid; Amino Acid Sequence; Animals; ATP; Catalysis; Cercopithecus aethiops; Chemical Biology; COS Cells; Glutamine/chemistry; Humans; Inbred C57BL; Leucine/chemistry; Lysine/chemistry; Mice; Molecular Sequence Data; Neutrophils/metabolism; Phenylalanine/chemistry; Phosphorylation; Protein Binding; Protein Kinase C (PKC); Protein Kinase C-delta/*metabolism; Protein Phosphorylation; Purines/chemistry; Sequence Homology; Signal Transduction; Stroke; Substrate Specificity; Superoxides/chemistry; Transgenic
To better study the role of PKCdelta in normal function and disease, we developed an ATP analog-specific (AS) PKCdelta that is sensitive to specific kinase inhibitors and can be used to identify PKCdelta substrates. AS PKCdelta showed nearly 200 times higher affinity (Km) and 150 times higher efficiency (kcat/Km) than wild type (WT) PKCdelta toward N(6)-(benzyl)-ATP. AS PKCdelta was uniquely inhibited by 1-(tert-butyl)-3-(1-naphthyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1NA-PP1) and
Kumar Varun; Weng Yi-Chinn; Geldenhuys Werner J; Wang Dan; Han Xiqian; Messing Robert O; Chou Wen-Hai
The Journal of biological chemistry
2015
2015-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1074/jbc.M114.598698" target="_blank" rel="noreferrer noopener">10.1074/jbc.M114.598698</a>
Cloning and characterization of two major 3-methylcholanthrene inducible hamster liver cytochrome P450s.
Female; Humans; Male; Animals; Rats; Amino Acid Sequence; Reference Values; Base Sequence; Liver/enzymology; Immunoblotting; Molecular Sequence Data; Cricetinae; Mesocricetus; Phenobarbital/pharmacology; Restriction Mapping; Nucleic Acid Hybridization; Cytochrome P-450 Enzyme System/biosynthesis/*genetics; Enzyme Induction; Liver/drug effects/*enzymology; Methylcholanthrene/*pharmacology; Inbred Strains; Sequence Homology; Cloning; Molecular; Nucleic Acid; Microsomes
We have studied the immunochemical properties of two major 3-methylcholanthrene inducible hamster liver cytochrome P450 isozymes, P450 MC1 and P450 MC4. Immunoblots using specific antibodies against P450 MC1 and P450 MC4 demonstrated that these two P450s were present in very low levels in control hamster livers and were greatly induced by 3-methylcholanthrene treatment. P450 MC1 was immunochemically different from P450 MC4, rat P450c and P450d, and rabbit LM4. The immunorelated polypeptide to P450 MC1 was not present in the control or the
Lai T S; Chiang J Y
Archives of biochemistry and biophysics
1990
1990-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/0003-9861(90)90664-k" target="_blank" rel="noreferrer noopener">10.1016/0003-9861(90)90664-k</a>
The expression of a catalytically active cholesterol 7 alpha-hydroxylase cytochrome P450 in Escherichia coli.
Gene Expression; Amino Acid Sequence; Base Sequence; Polymerase Chain Reaction; Liver/enzymology; Catalysis; Molecular Sequence Data; Substrate Specificity; Cholesterol/metabolism; Cholesterol 7-alpha-Hydroxylase/*genetics; DNA/genetics; Codon; Escherichia coli/*enzymology; Plasmids; Chromatography; Blotting; Western; Electrophoresis; Polyacrylamide Gel; Liquid; Microsomes
We have recently cloned a full-length cDNA encoding the rat hepatic cholesterol 7 alpha-hydroxylase cytochrome P450 (P450c7) (Li, Y. C., Wang, D. P., and Chiang, J. Y. L. (1990) J. Biol. Chem. 265, 12012-12019), which catalyzes the rate-limiting reaction of bile acid synthesis in the liver. By using the polymerase chain reaction, we have designed two P450c7 cDNAs. One has the second Met codon deleted and the third Thr codon replaced with an Ala. The other lacks codons for the NH2-terminal hydrophobic sequence of amino acids 2-24 (P450c7 delta 2-24). The cDNAs were separately cloned into the expression vector pKK233-2 and transformed into Escherichia coli. After induction with isopropyl-beta-D-thiogalactopyranoside, bacteria harboring recombinant plasmids expressed a polypeptide which reacted with the antibody against cholesterol 7 alpha-hydroxylase in immunoblots. The slightly modified full-length enzyme was expressed to 0.2% of the total bacterial lysate and was located in the membrane fraction, whereas P450c7 delta 2-24 was expressed at a 10-fold higher level (2%), of which 85% was in the cytosol and the remaining associated with the membranes. We have purified P450c7 delta 2-24 which showed a typical reduced-CO difference spectrum of cytochrome P450 and reconstituted cholesterol 7 alpha-hydroxylase activity in the presence of NADPH-cytochrome P450 reductase. P450c7 delta 2-24 has a similar Km for cholesterol (24.6 microM) but a lower Vmax (0.10 nmol/min) and a lower turnover number (1.93 min-1) as compared with the enzyme isolated from rat liver microsomes. The purified P450c7 delta 2-24 has an unique hydrophilic NH2 terminus and contains monomers and dimers in equal amounts. This is the first report demonstrating that a genetically engineered cytochrome P450 enzyme lacking a typical NH2-terminal hydrophobic sequence is mainly cytosolic and catalytically active.
Li Y C; Chiang J Y
The Journal of biological chemistry
1991
1991-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
Regulation of cholesterol 7 alpha-hydroxylase in the liver. Cloning, sequencing, and regulation of cholesterol 7 alpha-hydroxylase mRNA.
Female; Animals; Rats; Amino Acid Sequence; *Gene Expression Regulation; Kinetics; Base Sequence; Immunoblotting; Molecular Sequence Data; Steroid Hydroxylases/*genetics; Circadian Rhythm; DNA/genetics/isolation & purification; Restriction Mapping; Enzyme Induction; Liver/drug effects/*enzymology; Cell Fractionation; Cholesterol 7-alpha-Hydroxylase/biosynthesis/*genetics/immunology; Cholestyramine Resin/pharmacology; Cytochrome P-450 Enzyme System/genetics; Epitopes/analysis; Polyribosomes/metabolism/ultrastructure; Inbred Strains; RNA; Enzymologic; Sequence Homology; Cloning; Nucleic Acid; Messenger/*genetics; Centrifugation; Density Gradient; Molecular/methods
Monospecific antibody against purified rat liver cholesterol 7 alpha-hydroxylase cytochrome P-450 was used to screen a lambda gt11 cDNA library constructed from immuno-enriched polysomal RNA of cholestyramine-treated female rat liver. Two types of cDNA clones differing in the length of the 3'-untranslated region were identified, and DNA sequences were determined. The full length clone contains 3561 base pairs plus a long poly(A) tail. The amino acid sequence deduced from the open reading frame revealed a unique P-450 protein containing 503 amino acid residues which belonged to a new gene family designated family VII or CYP7. Southern blot hybridization experiments indicated that the minimal size of P-450 VII gene was 11 kilobase pairs (kb), and there was probably only one gene in this new family. Northern blot hybridization using specific cDNA probes revealed at least two major mRNA species of about 4.0 kb and 2.1 kb, respectively. These two mRNA species may be derived from the use of different polyadenylation signals and reverse-transcribed to two types of cDNA clones. Cholesterol 7 alpha-hydroxylase mRNAs were induced 2- to 3-fold in rat liver by cholestyramine treatment. The mRNA level was rapidly reduced upon the removal of the inducer. Similarly, cholesterol feeding induced enzyme activity, protein, and mRNA levels in the rat by 2-fold, suggesting that cholesterol is an important regulator of cholesterol 7 alpha-hydroxylase in the liver. On the other hand, dexamethasone and pregnenolone-16 alpha-carbonitrile drastically reduced the activity, protein, and mRNA levels. These experiments suggest that the induction of cholesterol 7 alpha-hydroxylase activity by cholestyramine or cholesterol and inhibition of cholesterol 7 alpha-hydroxylase activity by bile acid feedback are results of the rapid turnover of cholesterol 7 alpha-hydroxylase enzyme and mRNA levels.
Li Y C; Wang D P; Chiang J Y
The Journal of biological chemistry
1990
1990-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
An N-terminal arginine-rich cluster and a proline-alanine-threonine repeat region determine the cellular localization of the herpes simplex virus type 1 ICP34.5 protein and its ligand, protein phosphatase 1.
*Repetitive Sequences; Alanine/metabolism; Amino Acid; Amino Acid Sequence; Animals; Arginine/metabolism; Base Sequence; Cell Compartmentation; Cercopithecus aethiops; DNA Primers; Fluorescent Antibody Technique; Indirect; Ligands; Molecular Sequence Data; Phosphoprotein Phosphatases/*metabolism; Proline/metabolism; Protein Phosphatase 1; Recombinant Proteins/chemistry/metabolism; Subcellular Fractions/metabolism; Threonine/metabolism; Vero Cells; Viral Proteins/chemistry/*metabolism
The ICP34.5 protein facilitates herpes simplex virus replication by binding and activating protein phosphatase 1 (PP1) by means of a very conserved C-terminal
Mao Hanwen; Rosenthal Kenneth S
The Journal of biological chemistry
2002
2002-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1074/jbc.M111553200" target="_blank" rel="noreferrer noopener">10.1074/jbc.M111553200</a>
Tachykinin peptide-induced activation and desensitization of neurokinin 1 receptors.
Amino Acid Sequence; Animals; CHO Cells; Cricetinae; Down-Regulation/*drug effects; Electrophysiology; Ligands; Neurokinin-1/*agonists/*metabolism; Potassium/metabolism; Protein Binding; Rana catesbeiana; Receptors; Tachykinins/chemistry/*pharmacology
The actions of four tachykinins on inhibition and desensitization of the
Perrine S A; Bennett V J; Simmons M A
Peptides
2003
2003-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/s0196-9781(03)00064-0" target="_blank" rel="noreferrer noopener">10.1016/s0196-9781(03)00064-0</a>
The role of the N-terminal and mid-region residues of substance P in regulating functional selectivity at the tachykinin NK1 receptor.
Amino Acid Sequence; Animals; Binding; Calcium Signaling/physiology; CHO Cells; Competitive/drug effects; Cricetinae; Cricetulus; Dose-Response Relationship; Drug; Iodine Radioisotopes; Ligands; Models; Molecular; Molecular Sequence Data; Neurokinin-1/chemistry/drug effects/*metabolism; Rats; Receptors; Substance P/chemistry/drug effects/*physiology
Previous studies have shown that tachykinin peptide ligands of the tachykinin NK1 receptor exhibit functional selectivity with respect to signal activation and desensitization. The differences are most dramatic between the naturally occurring peptides substance P (RPKPQQFFGLM-NH2) and ranatachykinin C (HNPASFIGLM-NH2). To understand the structural features of the peptides that underlie these differences, four peptide analogs have been designed and tested. The analogs were designed to assess the major structural differences between substance P and ranatachykinin C, including the role of the N-terminal Arg and the substitution of the mid-region Glns with Ala and Ser (Q5 replaced with A and/or Q6 replaced with S). Receptor binding, receptor activation of intracellular calcium fluxes, and receptor desensitization of the rat tachykinin NK1 receptor were quantified for each ligand. All of the peptides bound to the rat tachykinin NK1 receptor with high affinity, produced robust calcium signal activation, and led to agonist-induced receptor desensitization. It was found that deletion of the N-terminal Arg of substance P or replacement of either or both Q5 and Q6 altered the functional selectivity of substance P based on the relationship of receptor binding to receptor activation and activation to desensitization. When considered in light of our previously published nuclear magnetic resonance structure data, the data presented herein suggest that the one, five and six positions of the substance P backbone are key structural residues that govern the relative degree of tachykinin peptide-mediated receptor signaling and desensitization.
Perrine Shane A; Beard Debbie J; Young John K; Simmons Mark A
European journal of pharmacology
2008
2008-09
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ejphar.2008.06.097" target="_blank" rel="noreferrer noopener">10.1016/j.ejphar.2008.06.097</a>
Immunization with a LEAPS heteroconjugate containing a CTL epitope and a peptide from beta-2-microglobulin elicits a protective and DTH response to herpes simplex virus type 1.
*Epitopes; Amino Acid Sequence; Animals; beta 2-Microglobulin/immunology; Conjugate/immunology; Cytotoxic/*immunology; Delayed/*etiology; Female; Herpesvirus 1; Human/*immunology; Hypersensitivity; Immediate-Early Proteins/*immunology; Immunization; Inbred BALB C; Mice; Molecular Sequence Data; Peptide Fragments/*immunology; T-Lymphocyte; T-Lymphocytes; Vaccines; Viral Vaccines/*immunology
A ligand epitope antigen presentation system (LEAPS) heteroconjugate vaccine containing a CTL epitope (H1) from the HSV-1 immediate early protein ICP27 (322-332) and a peptide sequence (J) from beta-2-microglobulin (35-50) elicited protection from intraperitoneal viral challenge and promoted DTH responses. The H1 peptide and other H1 containing heteroconjugates did not elicit protection or DTH responses. Antibody to the H1 peptide could not be detected by ELISA following vaccination with peptide, heteroconjugate or natural infection. The LEAPS heteroconjugate appears to prime a Thl-like response which is subsequently boosted by infection. These studies show that attachment of the J peptide can make a CTL epitope into a vaccine which is immunogenic and promotes a protective Th1 type of response.
Rosenthal K S; Mao H; Horne W I; Wright C; Zimmerman D
Vaccine
1999
1999-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/s0264-410x(98)00231-x" target="_blank" rel="noreferrer noopener">10.1016/s0264-410x(98)00231-x</a>
Induction of ethanol dependence increases signal peptidase mRNA levels in rat brain.
*Membrane Proteins; *Serine Endopeptidases; Alcoholism/*enzymology; Amino Acid; Amino Acid Sequence; Animals; Base Sequence; Blotting; Brain/*enzymology; Complementary; DNA; Endopeptidases/*biosynthesis/chemistry/genetics; Ethanol/*pharmacology; Male; Messenger/genetics/metabolism; Molecular Sequence Data; Northern; Nucleic Acid; Rats; RNA; Sequence Homology; Sprague-Dawley; Substance Withdrawal Syndrome/enzymology
Differential Northern blot hybridization was used as a screening tool to identify mRNAs that respond quantitatively to the induction of ethanol dependence. Adult male rats were treated with repeated, high doses of ethanol for 4 consecutive days. This regimen resulted in the development of tolerance and dependence upon ethanol. RNA isolated from the ethanol-dependent rat brains was used to construct a cDNA library. One cDNA was identified that hybridized to a mRNA which increased in rat brain during the ethanol treatment. Sequence analysis of the cDNA indicated that it recognized a mRNA in rat brain which was very similar to that which encodes the 18 kDa subunit of canine signal peptidase. The rat signal peptidase mRNA was observed to increase in brain nearly 2-fold within 48 h after the initiation of ethanol treatment. Ethanol did not significantly alter beta-actin mRNA levels during the treatment period. These results support the existence of an ethanol-responsive signal peptidase mRNA in rat brain.
Signs S A; Jacquet R
Molecular and cellular biochemistry
1994
1994-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/bf00944199" target="_blank" rel="noreferrer noopener">10.1007/bf00944199</a>
Expression of isotocin-neurophysin mRNA in developing zebrafish.
Amino Acid Sequence; Animals; Messenger/*metabolism; Molecular Sequence Data; Neurophysins/biosynthesis/*genetics; Oxytocin/*analogs & derivatives/biosynthesis/*genetics; Phylogeny; RNA; Zebrafish/*embryology
Neurohypophysial peptides are important regulators of homeostasis, reproduction and behavior. We have sequenced a zebrafish cDNA representing isotocin-neurophysin (IT-NP) mRNA. The developmental expression pattern of zebrafish IT-NP mRNA was determined by whole-mount in situ hybridization histochemistry. At 32 h post fertilization (hpf) no IT-NP mRNA is detected. However, by 36 hpf, staining for IT-NP mRNA is detected in a tight bilateral cluster of cells located in the anterior hypothalamus. The IT-NP mRNA expression pattern remains remarkably stable throughout further development at least until 120 hpf.
Unger Jennifer L; Glasgow Eric
Gene expression patterns : GEP
2003
2003-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/s1567-133x(02)00064-9" target="_blank" rel="noreferrer noopener">10.1016/s1567-133x(02)00064-9</a>
The isolation and characterization of the soluble and membrane-bound porcine cytochrome b5 cDNAs.
Amino Acid Sequence; Animals; Base Sequence; Blotting; Complementary/blood/*chemistry/*isolation & purification; Cytochromes b5/blood/*chemistry/genetics/*isolation & purification; DNA; Male; Membrane Proteins/*chemistry/genetics/*isolation & purification; Molecular Sequence Data; Solubility; Southern; Swine; Testis/enzymology
In some male pigs, there is an increased production of the testicular 16 androstene steroids which end up being concentrated in fatty tissue. When the meat is cooked, a disagreeable odor/flavor is produced, a phenomenon known as "boar taint." All boars selected for food production are castrated even though only ca 10% of boars may be "tainted." This has a high economic cost because castrated pigs convert food into meat less efficiently, the meat is fatter, and there is an increased mortality due to the castration procedure. Recent data has implicated an increased level of cytochrome b5 in the testes with the increased synthesis of the 16-androstene steroids. As an initial step in analyzing this process, we used 5' and 3' RACE PCR procedures to isolate, clone and sequence the cDNAs for the membrane-bound and soluble forms of porcine cytochrome b5.
VanDerMark P K; Steggles A W
Biochemical and biophysical research communications
1997
1997-11
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1006/bbrc.1997.7599" target="_blank" rel="noreferrer noopener">10.1006/bbrc.1997.7599</a>
Structure and nucleotide sequences of the human cholesterol 7 alpha-hydroxylase gene (CYP7).
Amino Acid Sequence; Base Sequence; Cholesterol 7-alpha-Hydroxylase/chemistry/*genetics; DNA; Humans; Molecular Sequence Data; Restriction Mapping
The human cholesterol 7 alpha-hydroxylase gene (CYP7) spans about 11 kb of the genome and contains six exons and five introns. Nucleotide sequences of a 5'-upstream region to exon III (5535 bp), a 5'-upstream EcoRI fragment (2575 bp), and an EcoRI fragment covering intron V to exon VI (2319 bp) have been determined. A comparison of our sequences with those reported previously unveiled numerous sequencing discrepancies that are apparently due to sequencing errors. There are only one confirmed and one possible genetic polymorphism in the promoter of this highly conserved human gene. The proximal promoter contained many consensus recognition sequences for liver-enriched transcription factors and steroid hormone receptors that may play important roles in regulation of the CYP7 gene transcription by bile acids, cholesterol, and hormones.
Wang D P; Chiang J Y
Genomics
1994
1994-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1006/geno.1994.1177" target="_blank" rel="noreferrer noopener">10.1006/geno.1994.1177</a>
Analysis of the thymidine kinase of a herpes simplex virus type 1 isolate that exhibits resistance to (E)-5-(2-bromovinyl)-2'-deoxyuridine.
Acyclovir/pharmacology; Amino Acid Sequence; Antiviral Agents/metabolism/*pharmacology; Base Sequence; Binding Sites; Bromodeoxyuridine/*analogs & derivatives/metabolism/pharmacology; Deoxyuridine/analogs & derivatives/pharmacology; DNA; Drug Resistance; Herpesvirus 1; Human/drug effects/*enzymology/genetics; Humans; Kinetics; Microbial; Molecular Sequence Data; Phosphonoacetic Acid/pharmacology; Phosphorylation; Point Mutation/genetics; Sequence Analysis; Thymidine Kinase/genetics/metabolism; Thymidine/metabolism; Vidarabine/pharmacology
The mechanism responsible for the decreased sensitivity of a clinical herpes simplex virus type 1 (HSV-1) isolate, HSV-145, to (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) was examined. Measurements of 50% inhibitory doses of several drugs demonstrated that although HSV-145 was sensitive to phosphonoacetic acid, adenine arabinoside and acyclovir, its sensitivity to BVDU and 5-(2-chloroethyl)-2'-deoxyuridine was significantly less than that normally observed for HSV-1. Analysis of the thymidylate kinase (TMP-K) activity of HSV-145 thymidine kinase (TK) demonstrated a decreased level of TMP-K activity when compared to HSV-1 TK. The TMP-K activity of HSV-145 resembled that observed for HSV-2 and the TK-deficient strain HSV-1 TK-7. When the nucleotide sequence of the HSV-145 TK gene was compared to that of the HSV-1 strains C1(101) and SC16 a single nucleotide substitution (G changed to A at base position 502) was detected which would result in the substitution of threonine at amino acid position 168 for alanine. The substitution is the same as that for the laboratory-derived BVDU-resistant virus HSV-1 SC16B3. Collectively, these studies highlight the importance of amino acid conservation at position 168 of the HSV-1 TK in conferring efficient TMP-K activity and BVDU sensitivity.
Wilber B A; Docherty J J
The Journal of general virology
1994
1994-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1099/0022-1317-75-7-1743" target="_blank" rel="noreferrer noopener">10.1099/0022-1317-75-7-1743</a>
The characterization of three types of partially processed mRNA and two pseudogenes for human liver cytochrome b5.
Humans; Amino Acid Sequence; Base Sequence; Molecular Sequence Data; Liver/*physiology; DNA/genetics; Cytochrome b Group/*genetics; Cytochromes b5; Pseudogenes; Cloning; Molecular; Post-Transcriptional; RNA Processing
We have isolated cDNA clones corresponding to partially processed human liver cytochrome b5 mRNAs. All the clones contained poly(A) sequences, and one clone had a shorter 3' non-translated sequence, indicating the use of an alternative poly(A) addition signal. In addition, all the clones contained the coding information for amino acids 87-134; however, there were two types of intron junction adjacent to the coding sequence. Detailed analysis of the Type I clones showed that the Type II intron sequence was contained within the Type I sequence, but approximately 1000 bp 5' of the Type I intron-exon junction showed alternative splicing within this intron. In addition, we have isolated two pseudogenes which lack introns, suggesting the retroviral insertion of human liver cytochrome b5 mRNA sequences into the human genome.
Yoo M; Steggles A W
Biochemical and biophysical research communications
1989
1989-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/0006-291x(89)92092-5" target="_blank" rel="noreferrer noopener">10.1016/0006-291x(89)92092-5</a>
The complete nucleotide sequence of human liver cytochrome b5 mRNA.
Humans; Amino Acid Sequence; Liver/*metabolism; Base Sequence; Molecular Sequence Data; DNA/genetics; Cytochrome b Group/*genetics; Cytochromes b5; RNA; Cloning; Molecular; Messenger/*genetics
We have isolated and sequenced a cDNA clone corresponding to human liver cytochrome b5 mRNA. The 760 base pair (bp) sequence contains the complete coding and 3' non-translated regions plus 52 bp of 5' non-translated sequence. The derived amino acid sequence showed that the previous assignment of several amino acids was in error. In addition, the sequence of the previously unknown COOH hydrophobic region has been obtained.
Yoo M; Steggles A W
Biochemical and biophysical research communications
1988
1988-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/s0006-291x(88)80881-7" target="_blank" rel="noreferrer noopener">10.1016/s0006-291x(88)80881-7</a>
Promoter activity and regulation of the CYP4F2 leukotriene B(4) omega-hydroxylase gene by peroxisomal proliferators and retinoic acid in HepG2 cells.
*Gene Expression Regulation; *Promoter Regions; Amino Acid Sequence; Base Sequence; Cell Line; Cloning; Cytochrome P-450 CYP4A; Cytochrome P-450 Enzyme System/*genetics/metabolism; Cytochrome P450 Family 4; Cytoplasmic and Nuclear/metabolism; DNA; DNA Footprinting; Enzymologic; Exons; Genes; Genetic; Genetic/drug effects; Humans; Introns; Leukotriene B4/metabolism; Mixed Function Oxygenases/metabolism; Models; Molecular; Molecular Sequence Data; Peroxisome Proliferators/*metabolism; Receptors; Reporter; Retinoic Acid Receptor alpha; Retinoic Acid/metabolism; Sequence Analysis; Transcription; Transcription Factors/metabolism; Transfection; Tretinoin/*metabolism
The human liver CYP4F2 gene (Accession No. AF221943) encodes a leukotriene B(4) omega-hydroxylase that metabolizes leukotriene B(4) (LTB(4)) to a less potent proinflammatory eicosanoid, 20-OH-LTB(4). We sequenced a 6.7-kb genomic fragment of the human CYP4F2 gene that has the first five exons and 500 bp of the 5'-flanking region. The major transcription start site was found to be 49 bp upstream of the 3' end of exon 1 and the ATG translation initiation codon was located in exon 2. Besides the TATA box at -39 bp and basal transcription factor binding sites, the promoter region and 412-bp intron 1 have several putative binding sites for nuclear factors that may mediate the inflammatory response and lipid homeostasis. We found two DR1 elements in the 5' promoter, a DR2 element in intron 1, and RXR/RAR binding sites in both intron 1 and the 5' promoter. DNase I footprinting revealed three protected sequences, with the region containing two CAATT boxes at -71 and -111 bp important in CYP4F2 gene expression. Luciferase reporter assays showed that the 500-bp upstream sequence has strong promoter activity. Transient transfection experiments identified two sites in the 5' promoter and intron 1 that cooperate in gene transcription while exon 1 and a
Zhang X; Chen L; Hardwick J P
Archives of biochemistry and biophysics
2000
2000-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1006/abbi.2000.1836" target="_blank" rel="noreferrer noopener">10.1006/abbi.2000.1836</a>