title: Methylation and carbamylation of human gamma-crystallins.
abstract: Accessible sulfhydryls of cysteine residues are likely sites of reaction in long-lived proteins such as human lens crystallins. Disulfide bonding between cysteines is a major contributor to intermolecular cross-linking and aggregation of crystallins. A recently reported modification of gammaS-crystallins, S-methylation of cysteine residues, can prevent disulfide formation. The aim of this study was to determine whether cysteines in gammaC-, gammaD-, and gammaB-crystallins are also S-methylated. Our data show that all the gamma-crystallins are S-methylated, but only at specific cysteines. In gammaD-crystallin, methylation is exclusively at Cys 110, whereas in gammaC- and gammaB-crystallins, the principal methylation site is Cys 22 with minor methylation at Cys 79. gammaD-crystallin is the most heavily methylated gamma-crystallin. gammaD-Crystallins from adult lenses are 37%-70% methylated, whereas gammaC and gammaB are approximately 12% methylated. The specificity of gamma-crystallin methylation and its occurrence in young clear lenses supports the idea that inhibition of disulfide bonding by S-methylation may play a protective role against cataract. Another modification, not reported previously, is carbamylation of the N termini of gammaB-, gammaC-, gammaD-crystallins. N-terminal carbamylation is likely a developmentally related modification that does not negatively impact crystallin function.
authors.affiliation: Department of Chemistry, University of Nebraska, Lincoln, NE 68588, USA.
authors.forename: Veniamin N
authors.initials: VN
authors.lastname: Lapko
authors.forename: David L
authors.initials: DL
authors.lastname: Smith
authors.forename: Jean B
authors.initials: JB
authors.lastname: Smith
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000502:physiology; D000595:Amino Acid Sequence; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007908:Lens, Crystalline / Q000737:chemistry / Q000378:metabolism*; D008745:Methylation; D008875:Middle Aged; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D017386:Sequence Homology, Amino Acid; D021241:Spectrometry, Mass, Electrospray Ionization; D014357:Trypsin / Q000378:metabolism; D038222:gamma-Crystallins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D038222:gamma-Crystallins; D014357:Trypsin