S-methylated cysteines in human lens gamma S-crystallins.
The proteins of the eye lens, which do not turn over throughout life, undergo many modifications, some of which lead to senile cataract. We describe a modification, S-methylation of cysteine, that may serve to protect the lens from detrimental modifications. The modification was detected as a +14 Da peak in electrospray ionization mass spectra of human lens gammaS-crystallins. Derivatization of gammaS-crystallin with iodoacetamide showed reaction at only six of the seven cysteines, indicating the modification blocked reaction at one cysteine. Further analysis of the modified gammaS-crystallin as tryptic peptides located the modification primarily at Cys 26, with smaller amounts at Cys 24. Tandem mass spectrometry and exact mass measurements showed that the modification was S-methylation. Methylation of proteins has been documented at several other amino acid residues, but S-methylation of cysteine residues has previously been detected only as part of a methyltransferase DNA repair mechanism or at trace amounts in hemoglobin. The high levels of S-methylated cysteines in lens nuclei and the specificity for Cys 26 and Cys 24 suggest the reaction is enzymatically mediated. This modification is particularly important because it blocks disulfide bonding of gammaS-crystallins and, thereby, inhibits formation of the high-molecular weight assemblies associated with cataract. Evidence of more S-methylation in soluble than in insoluble gammaS-crystallins supports the contention that S-methylation of gammaS-crystallin inhibits protein insolubilization and may offer protection against cataract.
Lapko
Veniamin N
VN
Department of Chemistry, University of Nebraska, Lincoln 68588-0304, USA.
Smith
David L
DL
Smith
Jean B
JB
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000378:metabolism; D002918:Chymotrypsin / Q000737:chemistry; D003545:Cysteine / Q000032:analysis / Q000737:chemistry*; D006801:Humans; D007231:Infant, Newborn; D007908:Lens, Crystalline / Q000737:chemistry*; D008745:Methylation; D008875:Middle Aged; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D038222:gamma-Crystallins / Q000737:chemistry* / Q000302:isolation & purification
D010446:Peptide Fragments; D013438:Sulfhydryl Compounds; D038222:gamma-Crystallins; C077648:CRYGS protein, human; D002918:Chymotrypsin; D003545:Cysteine