title: Age-related changes in bovine alpha-crystallin and high-molecular-weight protein.
abstract: The high-molecular-weight (HMW) protein from the lens is composed mostly of alpha-crystallin in a highly aggregated state. Bovine HMW protein was carefully separated from alpha-crystallin by size-exclusion chromatography. alpha-Crystallin has chaperone-like ability whereby it stabilizes other proteins under conditions of stress (e.g. heat). Comparison of bovine HMW protein and alpha-crystallin shows that the HMW protein has a markedly reduced chaperone ability compared to alpha-crystallin. However, in contrast to the results of other workers, we observe no alteration with age in the ability of alpha-crystallin to act as a chaperone. Using electrospray ionisation mass spectrometry, changes in the phosphorylation of the alpha-crystallin subunits with age have been quantified. Phosphorylation of alpha-crystallin occurs early in life but does not alter in proportion after about three years of age. In addition, phosphorylation of the A subunit of alpha-crystallin has little effect on its chaperone ability. As is found in the artificially prepared HMW complex of alpha- and gamma-crystallin, NMR spectroscopy shows that in the naturally occurring HMW protein, the short C-terminal extension of the alpha B subunit has lost its flexibility whereas the alpha A subunit extension is still flexible. Post-translational modifications therefore seem to have little effect on the chaperone action of alpha-crystallin, but alterations in the quaternary structure of alpha-crystallin via incorporation into the HMW aggregate, lead to major changes in the chaperone ability of the protein. The results are consistent with the notion that one of the contributing factors to cataract formation in the lens is the depletion of alpha-crystallin with age as it is converted into the HMW protein.
authors.affiliation: Australian Cataract Research Foundation, Department of Chemistry, University of Wollongong, NSW, Australia.
authors.forename: J A
authors.initials: JA
authors.lastname: Carver
authors.forename: K A
authors.initials: KA
authors.lastname: Nicholls
authors.forename: J A
authors.initials: JA
authors.lastname: Aquilina
authors.forename: R J
authors.initials: RJ
authors.lastname: Truscott
mesh_terms: D000375:Aging / Q000378:metabolism*; D000426:Alcohol Dehydrogenase / Q000737:chemistry; D000818:Animals; D002417:Cattle; D003459:Crystallins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D007908:Lens, Crystalline / Q000737:chemistry* / Q000502:physiology; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008970:Molecular Weight; D010766:Phosphorylation; D013329:Structure-Activity Relationship
chemical_list: D003459:Crystallins; D000426:Alcohol Dehydrogenase