title: Identification of multiple metabolic enzymes from mice cochleae tissue using a novel functional proteomics technology.
abstract: 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.
authors: Department of Biology, Vanderbilt University, Nashville, TN, United States of America.
authors: David L
authors: DL
authors: Wang
authors: Department of Otolaryngology, School of Medicine, Washington University, St. Louis, MO, United States of America.
authors: Hui
authors: H
authors: Li
authors: Department of Otolaryngology, School of Medicine, Washington University, St. Louis, MO, United States of America.
authors: Ruqiang
authors: R
authors: Liang
authors: Department of Otolaryngology, School of Medicine, Washington University, St. Louis, MO, United States of America.
authors: Jianxin
authors: J
authors: Bao
mesh_terms: D000818:Animals; D002417:Cattle; D003051:Cochlea / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D009097:Multienzyme Complexes / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism; D010084:Oxidation-Reduction; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D000069466:Red Meat
chemical_list: D009097:Multienzyme Complexes; D020543:Proteome; D009243:NAD; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases