Evidence for the participation of alpha B-crystallin in human age-related nuclear cataract.
The aim of this study was to determine if the unusual coloured species characteristic of age-related nuclear cataract could be localised to specific residues of the crystallins. The insoluble, crosslinked and coloured cataract protein fraction (CPF) was isolated from cataract human lenses. Using a combination of tryptic digestion, gel filtration and multiple reversed phase high performance liquid chromatography (RP-HPLC), coloured peaks were isolated and subjected to amino acid sequence analysis. With these techniques, it was hoped to identify and locate the modified residues. Sequence information was obtained on 16 'coloured' peptides. Many of the peptides were found to be derived from alpha B-crystallin. When redundancies are taken into account, six distinctive peptides were found to be derived from alpha B-crystallin; one from beta B1-crystallin, two from beta A3/A1-crystallin and three from gamma S-crystallin. Three sites of possible crystallin residue isomerisation to modification were detected in the alpha B- and beta A3/beta A1-crystallins, including probable asp isomerisation at residues 25 and 36 in alpha B-crystallin. Since the CPF is unique to nuclear cataract lenses, these data suggest that alpha-crystallin, and alpha B-crystallin in particular, may be implicated in the cataract process. This finding supports that of a recent study on cataract proteins using pronase digestion [Chen YC, Reid GE, Simpson RJ, Truscott RJW. Exp Eye Res 1997;65:835.]
Truscott
R J
RJ
Australian Cataract Research Foundation, University of Wollongong, NSW, Australia.
Chen
Y C
YC
Shaw
D C
DC
D000375:Aging / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D002386:Cataract / Q000209:etiology* / Q000235:genetics / Q000378:metabolism*; D003116:Color; D003432:Cross-Linking Reagents; D003459:Crystallins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007907:Lens Nucleus, Crystalline / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012995:Solubility
D003432:Cross-Linking Reagents; D003459:Crystallins; D010446:Peptide Fragments