Post-translational modification of crystallins in vitreous body from experimental autoimmune uveitis of rats.
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 alphaB-crystallin. Thirty spots were identified in vitreous bodies, and 27 of these spots were members of the crystallin family. Unlike betaA4- and B2-crystallins (that were significantly increased without truncation), alphaA- 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 alphaA- and alphaB-crystallins, phosphorylation of alphaB-crystallin, and new production of betaA4- and betaB2-crystallins are intercorrelated with uveitis progression and inflammatory responses.
Bahk
Song-Chul
SC
Genome Research Center for Immune Disorders, School of Medicine, Wonkwang University, Iksan, Republic of Korea.
Jang
Jung-Un
JU
Choi
Chang-Uk
CU
Lee
Sook-Hee
SH
Park
Zee-Yong
ZY
Yang
Ji-Yeon
JY
Kim
Jae-Duck
JD
Yang
Yun-Sik
YS
Chung
Hun-Taeg
HT
D000818:Animals; D001327:Autoimmune Diseases / Q000378:metabolism*; D002853:Chromatography, Liquid; D003459:Crystallins / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006427:Hemin / Q000494:pharmacology; D008297:Male; D008665:Metalloporphyrins / Q000494:pharmacology; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011524:Protoporphyrins / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D015866:Uveitis, Posterior / Q000378:metabolism*; D014822:Vitreous Body / Q000378:metabolism*; D038202:alpha-Crystallin A Chain / Q000378:metabolism; D038203:alpha-Crystallin B Chain / Q000378:metabolism