Identification of Multiple Metabolic Enzymes from Mice Cochleae Tissue Using a Novel Functional Proteomics Technology

Identification of Multiple Metabolic Enzymes from Mice Cochleae Tissue Using a Novel Functional Proteomics Technology

Wang D L; Li H; Liang R Q; Bao J X

PLOS ONE

2015

2015-03

A new type of technology in proteomics was developed in order to separate a complex protein mixture and analyze protein functions systematically. The technology combines the ability of two-dimensional gel electrophoresis (2-DE) to separate proteins with a protein elution plate (PEP) to recover active proteins for functional analysis and mass spectrometry (MS)-based identification. In order to demonstrate the feasibility of this functional proteomics approach, NADH and NADPH-dependent oxidases, major redox enzyme families, were identified from mice cochlear tissue after a specific drug treatment. By comparing the enzymatic activity between mice that were treated with a drug and a control group significant changes were observed. Using MS, five NADH-dependent oxidases were identified that showed highly altered enzymatic activities due to the drug treatment. In essence, the PEP technology allows for a systematic analysis of a large enzyme family from a complex proteome, providing insights in understanding the mechanism of drug treatment.

Animal tissues; Beef; Biology; cancer; cancer metabolism; cell metabolism; Cochlea; draft; Drug therapy; E coli; Electrophoresis; Elution; Enzymatic activity; Enzyme assays; Enzyme metabolism; Enzymes; Feasibility studies; Functional analysis; Gel electrophoresis; Genes; Genomics; Hearing impairment; Kinases; Mass spectrometry; Mass spectroscopy; medicine; messenger-rna; metabolism; mice; NAD(P)H oxidase; NADH; Neurobiology; Neurosciences; Nicotinamide adenine dinucleotide; Noise; Otolaryngology; Oxidation-reduction reactions; Phosphatase; Protein expression; proteins; Proteomes; Proteomics; Science & Technology - Other Topics; Sciences: Comprehensive Works; Scientific imaging; Studies; Substance abuse treatment; Technology; United States--US

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