Post-translational modification of betaH-crystallin of bovine lens with aging.
This study investigates post-translational modification of proteins of bovine lens with aging (3 year old vs. 6 month old cows). After water-soluble proteins were submitted to gel and ion exchange chromatography, betaH-crystallin, a subunit of beta-crystallin, and modified materials were isolated. These materials were then submitted to two dimensional polyacrylamide gel electrophoresis (2D-SDS PAGE) to detect and isolate the new spots. Results for lens proteins from 3 year old animals were compared to those from 6 month old animals. All spots were digested in gel with trypsin and the molecular masses of tryptic digests were measured by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOFMS). Peptides peaks obtained from mass mapping were identified using the protein database of the MS-Fit program in the Protein prospector program of the University of California, San Francisco. We found that two post translational modifications of betaH-crystallin, acetylation and phosphorylation occurred with aging.
Kamei
Akira
A
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Meijo University, Nagoya, Aichi, Japan. kamei@ccmfs.meijo-u.ac.jp
Takeuchi
Noriko
N
Nagai
Makoto
M
Mori
Satoshi
S
D000107:Acetylation; D000375:Aging; D000818:Animals; D002140:California; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D003459:Crystallins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D007908:Lens, Crystalline / Q000737:chemistry* / Q000378:metabolism*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D012495:San Francisco; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014357:Trypsin / Q000378:metabolism
D003459:Crystallins; D014357:Trypsin