Supramolecular order within the lens: 1H NMR spectroscopic evidence for specific crystallin-crystallin interactions.
alpha-, beta- and gamma-crystallins from bovine lens contain flexible terminal extensions which are readily observed by NMR spectroscopy. By monitoring these resonances, NMR spectroscopy therefore offers a means of examining specific protein-protein interactions in crystallin mixtures. In this paper, a 1H NMR spectroscopic study of bovine lens nuclear and cortical homogenates and various crystallin mixtures is presented. In both homogenates, resonances from the flexible C-terminal extensions of alpha-crystallin and the N-terminal extension of beta B2-crystallin are readily observed suggesting that these regions are not involved in crystallin-crystallin interactions. In the cortical homogenate, resonances from the short N-terminal extension of gamma S-crystallin are also present. The cortical homogenate gives rise to more intense resonances than the nuclear homogenate, suggesting that the cortical region has many more mobile crystallin regions. In both homogenates, the C-terminal extension of beta B2-crystallin and the very short C-terminal extension of gamma B-crystallin are not observed. Thus, the C-terminal regions of these proteins are involved in interactions with other crystallins. Similar effects are observed upon mixing of the individual crystallins, e.g. the C-terminal extension of gamma B-crystallin is absent in spectra of mixtures of total gamma-crystallin and high-molecular-weight beta-crystallin aggregates (beta H). Overall, the results are consistent with a short-range order for the crystallins within the lens.
Cooper
P G
PG
Australian Cataract Research Foundation, Department of Chemistry, University of Wollongong, NSW, Australia.
Aquilina
J A
JA
Truscott
R J
RJ
Carver
J A
JA
D000818:Animals; D002417:Cattle; D003459:Crystallins / Q000737:chemistry*; D007904:Lens Cortex, Crystalline / Q000737:chemistry; D007907:Lens Nucleus, Crystalline / Q000737:chemistry; D007908:Lens, Crystalline / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy
D003459:Crystallins