Post-Translational Modification of Crystallins in Vitreous Body from Experimental Autoimmune Uveitis of Rats

• Published in: Journal of proteome research 2007-10, Vol.6 (10), p.3891-3898
• Main Authors: Bahk, Song-Chul, Jang, Jung-Un, Choi, Chang-Uk, Lee, Sook-Hee, Park, Zee-Yong, Yang, Ji-Yeon, Kim, Jae-Duck, Yang, Yun-Sik, Chung, Hun-Taeg
• Format: Article
• Language: English
• Subjects: alpha-Crystallin A Chain - metabolism; alpha-Crystallin B Chain - metabolism; Animals; Autoimmune Diseases - metabolism; Chromatography, Liquid; Crystallins - metabolism; Electrophoresis, Gel, Two-Dimensional; Hemin - pharmacology; Male; Metalloporphyrins - pharmacology; Phosphorylation; Protein Processing, Post-Translational; Protoporphyrins - pharmacology; Rats; Rats, Inbred Lew; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tandem Mass Spectrometry; Uveitis, Posterior - metabolism; Vitreous Body - metabolism
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/pr070133k

Experimental autoimmune uveitis (EAU) is a well-known animal model of posterior uveitis that is one of the major causes of blindness. EAU could be induced in susceptible animals (i.e., Lewis rat) by immune reactions using evolutionarily conserved retinal proteins, such as interphoto-receptor retinoid binding protein (IRBP), or epitaphs of the protein. First, we prepared the following four test groups that subsequently increased or decreased inflammation. (1) Normal control group, (2) IRBP-induced uveitis group, (3) Hemin-treated uveitis group, and (4) Sn(IV) protoporphyrin IX dichloride (SnPP)-treated uveitis group. Second, in the vitreous bodies of Lewis rats, the infiltrated proteins were analyzed using two-dimensional electrophoresis (2-DE), MALDI-TOF/MS, and Micro LC/LC−MS/MS analysis. Finally, Western blotting was applied to confirm the relative amount of crystallins and phosphorylation sites of αB-crystallin. Thirty spots were identified in vitreous bodies, and 27 of these spots were members of the crystallin family. Unlike βA4- and B2-crystallins (that were significantly increased without truncation), αA- and B-crystallins were only truncated in EAU vitreous body. Taken as a whole, in the rat EAU model, we suggest that post-translational truncations of αA- and αB-crystallins, phosphorylation of αB-crystallin, and new production of βA4- and βB2-crystallins are intercorrelated with uveitis progression and inflammatory responses. Keywords: crystallin • experimental autoimmune uveitis (EAU) • vitreous body • truncation • phosphorylation