Profiling of acyl-CoA oxidase-deficient and peroxisome proliferator Wy14,643-treated mouse liver protein by surface-enhanced laser desorption/ionization ProteinChip Biology System.
Peroxisome proliferators induce hepatic peroxisome proliferation and hepatocellular carcinomas in rodents. These chemicals increase the expression of the peroxisomal beta-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 beta-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 Wyl4,643-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.
Chu
Ruiyin
R
Department of Functional Genomics, Aventis Pharmaceuticals, Inc., Bridgewater, NJ 08807, USA. ruiyin.chu@aventis.com
Zhang
Weihua
W
Lim
Hanjo
H
Yeldandi
Anjana V
AV
Herring
Chris
C
Brumfield
Laura
L
Reddy
Janardan K
JK
Davison
Matthew
M
D042962:Acyl-CoA Oxidase; D000818:Animals; D002455:Cell Division; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010088:Oxidoreductases / Q000235:genetics* / Q000502:physiology; D010455:Peptides / Q000737:chemistry; D020025:Peroxisome Proliferators / Q000494:pharmacology*; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D011743:Pyrimidines / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D010455:Peptides; D020025:Peroxisome Proliferators; D011506:Proteins; D020543:Proteome; D011743:Pyrimidines; C035737:major urinary proteins; C006253:pirinixic acid; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase