title: Fluorescence resonance energy transfer study of subunit exchange in human lens crystallins and congenital cataract crystallin mutants.
abstract: Lens alpha-crystallin is an oligomeric protein with a molecular mass of 500-1000 kDa and a polydispersed assembly. It consists of two types of subunits, alphaA and alphaB, each with a molecular mass of 20 kDa. The subunits also form homo-oligomers in some other tissues and in vitro. Their quaternary structures, which are dynamic and characterized by subunit exchange, have been studied by many techniques, including fluorescence resonance energy transfer (FRET) and mass spectrometry analysis. The proposed mechanism of subunit exchange has been either by dissociation/association of monomeric subunits or by rapid equilibrium between oligomers and suboligomers. To explore the nature of subunit exchange further, we performed additional FRET measurements and analyses using a fluorescent dye-labeled W9F alphaA-crystallin as the acceptor probe and Trp in other crystallins (wild-type and R116C alphaA, wild-type and R120G alphaB, wild-type and Q155* betaB2) as the donor probe and calculated the transfer efficiency, Förster distance, and average distance between two probes. The results indicate only slight decreased efficiency and increased distance between two probes for the R116C alphaA and R120G alphaB mutations despite conformational changes.
authors.affiliation: Ophthalmic Research/Surgery, Brigham and Women's Hospital, Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA. jliang@rics.bwh.harvard.edu
authors.forename: Jack J
authors.initials: JJ
authors.lastname: Liang
authors.forename: Bing-Fen
authors.initials: BF
authors.lastname: Liu
mesh_terms: D002386:Cataract / Q000151:congenital / Q000209:etiology / Q000235:genetics*; D003459:Crystallins / Q000737:chemistry* / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007908:Lens, Crystalline / Q000737:chemistry*; D009154:Mutation; D021122:Protein Subunits / Q000378:metabolism; D013696:Temperature
chemical_list: D003459:Crystallins; D021122:Protein Subunits