Amyloid fibril formation by lens crystallin proteins and its implications for cataract formation.
The alpha-, beta-, and gamma-crystallins are the major structural proteins within the eye lens and are responsible for its exceptional stability and transparency. Under mildly denaturing conditions, all three types of bovine crystallin assemble into fibrillar structures in vitro. Characterization by transmission electron microscopy, dye binding assays, and x-ray fiber diffraction shows that these species have all of the characteristics of fibrils associated with the family of amyloid diseases. Moreover, the full-length proteins are incorporated into the fibrils, (i.e. no protein cleavage is required for these species to form), although for the gamma-crystallins some fragmentation occurs under the conditions employed in this study. Our findings indicate that the inherent stability of the beta-sheet supramolecular structure adopted by the crystallins in the eye lens and the chaperone ability of alpha-crystallin must be crucial for preventing fibril formation in vivo. The crystallins are very stable proteins but undergo extensive post-translational modification with age that leads to their destabilization. The ability of the crystallins to convert into fibrils under destabilizing conditions suggests that this process could contribute to the development of cataract with aging.
Meehan
Sarah
S
Biological Physics Group, Cavendish Laboratory, University of Cambridge, Madingley Road, Cambridge CB3 0HE, United Kingdom.
Berry
Yoke
Y
Luisi
Ben
B
Dobson
Christopher M
CM
Carver
John A
JA
MacPhee
Cait E
CE
D000375:Aging; D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D052160:Benzothiazoles; D002386:Cataract / Q000473:pathology*; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D019791:Guanidine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007908:Lens, Crystalline / Q000378:metabolism*; D008854:Microscopy, Electron; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D013696:Temperature; D013844:Thiazoles / Q000737:chemistry; D014961:X-Ray Diffraction; D038201:alpha-Crystallins / Q000737:chemistry*; D038204:beta-Crystallins / Q000737:chemistry*; D038222:gamma-Crystallins / Q000737:chemistry*
D000682:Amyloid; D052160:Benzothiazoles; D013844:Thiazoles; D038201:alpha-Crystallins; D038204:beta-Crystallins; D038222:gamma-Crystallins; C009462:thioflavin T; D019791:Guanidine