The oxidized thiol proteome in aging and cataractous mouse and human lens revealed by ICAT labeling.
Age-related cataractogenesis is associated with disulfide-linked high molecular weight (HMW) crystallin aggregates. We recently found that the lens crystallin disulfidome was evolutionarily conserved in human and glutathione-depleted mouse (LEGSKO) cataracts and that it could be mimicked by oxidation in vitro (Mol. Cell Proteomics, 14, 3211-23 (2015)). To obtain a comprehensive blueprint of the oxidized key regulatory and cytoskeletal proteins underlying cataractogenesis, we have now used the same approach to determine, in the same specimens, all the disulfide-forming noncrystallin proteins identified by ICAT proteomics. Seventy-four, 50, and 54 disulfide-forming proteins were identified in the human and mouse cataracts and the in vitro oxidation model, respectively, of which 17 were common to all three groups. Enzymes with oxidized cysteine at critical sites include GAPDH (hGAPDH, Cys247), glutathione synthase (hGSS, Cys294), aldehyde dehydrogenase (hALDH1A1, Cys126 and Cys186), sorbitol dehydrogenase (hSORD, Cys140, Cys165, and Cys179), and PARK7 (hPARK7, Cys46 and Cys53). Extensive oxidation was also present in lens-specific intermediate filament proteins, such as BFSP1 and BFSP12 (hBFSP1 and hBFSP12, Cys167, Cys65, and Cys326), vimentin (mVim, Cys328), and cytokeratins, as well as microfilament and microtubule filament proteins, such as tubulin and actins. While the biological impact of these modifications for lens physiology remains to be determined, many of these oxidation sites have already been associated with either impaired metabolism or cytoskeletal architecture, strongly suggesting that they have a pathogenic role in cataractogenesis. By extrapolation, these findings may be of broader significance for age- and disease-related dysfunctions associated with oxidant stress.
Center for Proteomics, Case Western Reserve University, Cleveland, OH, 44120, USA.
Benlian
B
Wang
Department of Pathology, Case Western Reserve University, Cleveland, OH, 44120, USA.
Grant
G
Hom
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Sheng
S
Zhou
Department of Ophthalmology, The Huichang County People's Hospital, Jiangxi, China.
Minfei
M
Guo
Department of Ophthalmology, Ganzhou City People's Hospital, Jiangxi, China.
Binbin
B
Li
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Jing
J
Yang
Department of Pathology, Case Western Reserve University, Cleveland, OH, 44120, USA.|Department of Biochemistry, Case Western Reserve University, Cleveland, OH, 44120, USA.
Vincent M
VM
Monnier
Department of Pathology, Case Western Reserve University, Cleveland, OH, 44120, USA.
Xingjun
X
Fan
D000293:Adolescent; D000368:Aged; D000375:Aging / Q000378:metabolism* / Q000473:pathology; D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D002386:Cataract / Q000378:metabolism* / Q000473:pathology; D002648:Child; D002675:Child, Preschool; D003432:Cross-Linking Reagents / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D007908:Lens, Crystalline / Q000378:metabolism* / Q000473:pathology; D018345:Mice, Knockout; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D000071617:Protein Deglycase DJ-1 / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D000975:Antioxidants; D003432:Cross-Linking Reagents; D010455:Peptides; D020543:Proteome; D013438:Sulfhydryl Compounds; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1; D005978:Glutathione; D003545:Cysteine
aging; cataractogenesis; disulfide; mass spectrometry; proteomics; reactive oxygen species