Profiling of Acyl-CoA Oxidase-Deficient and Peroxisome Proliferator Wy14,643-Treated Mouse Liver Protein by Surface-Enhanced Laser Desorption/Ionization ProteinChip® Biology System

• Published in: Gene expression 2002-01, Vol.10 (4), p.165-177
• Main Authors: Chu, Ruiyin, Zhang, Weihua, Lim, Hanjo, Yeldandi, Anjana V, Herring, Chris, Brumfield, Laura, Reddy, Janardan K, Davison, Matthew
• Format: Article
• Language: English
• Subjects: Acyl-CoA Oxidase; Animals; Cell Division; Chromatography, Liquid; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Liver - drug effects; Liver - metabolism; Male; Mass Spectrometry; Mice; Mice, Inbred C57BL; Mice, Transgenic; Oxidoreductases - genetics; Oxidoreductases - physiology; Peptides - chemistry; Peroxisome Proliferators - pharmacology; Protein Array Analysis - methods; Proteinchip Array Seldi Mass Spectrometry Peroxiso; Proteins - metabolism; Proteome - analysis; Proteomics; Pyrimidines - pharmacology; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 1052-2166; 1555-3884
• DOI: 10.3727/000000002783992460

Peroxisome proliferators induce hepatic peroxisome proliferation and hepatocellular carcinomas in rodents. These chemicals increase the expression of the peroxisomal β-oxidation pathway and the cytochrome P-450 4A family, which metabolizes lipids, including fatty acids. Mice lacking fatty acyl-CoA oxidase (AOX-/-), the first enzyme of the peroxisomal β-oxidation system, exhibit extensive microvesicular steatohepatitis, leading to hepatocellular regeneration and massive peroxisome proliferation. To investigate proteins involved in peroxisome proliferation, we adopted a novel surface-enhanced laser desorption/ionization (SELDI) ProteinChip technology to compare the protein profiles of control (wild-type), AOX-/-, and wild-type mice treated with peroxisome proliferator, Wy-14,643. The results indicated that the protein profiles of AOX-/- mice were similar to the wild-type mice treated with Wy14,643, but significantly different from the nontreated wild-type mice. Using four different ProteinChip Arrays, a total of 40 protein peaks showed more than twofold changes. Among these differentially expressed peaks, a downregulated peak was identified as the major urinary protein in both AOX-/- and Wy14,643-treated mice by SELDI. The identification of MUP was further confirmed by two-dimensional electrophoresis and liquid chromatography coupled tandem mass spectrometry (LC-MS-MS). This SELDI method offers several technical advantages for detection of differentially expressed proteins, including ease and speed of screening, no need for chromatographic processing, and small sample size.