title: The effect of temperature on the renaturation of alpha-crystallin.
abstract: The effects of variations in temperature and protein concentration on the renaturation of bovine alpha-crystallin have been examined using gel permeation chromatography, sedimentation analysis, fluorescence spectroscopy and electron microscopy. High protein concentration (3-53 mg/ml) were found to generate heterogenous populations of aggregates. It was concluded that concentrations above 3 mg/ml were inappropriate for renaturation of alpha-crystallin. Aggregates with molecular masses gradually increasing from 461,000 to 695,000 Da were produced with increasing temperature over the range 6-39 degrees C. Electron microscopy demonstrated that the reaggregates were composed predominantly of particles with circular cross-sections and mean diameters of 13-14 nm. As the renaturation temperature increased, increasing amounts of sheet-like structures were observed. Tryptophan accessibility to acrylamide quenching decreased in these aggregates as the size increased. These observations are consistent with the concept that there is no unique quaternary structure, or set of structures, for alpha-crystallin but that the protein can exist in a variety of forms containing different numbers of subunits.
authors.affiliation: Russell Grimwade School of Biochemistry, Parkville, Victoria, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Clauwaert
authors.forename: H D
authors.initials: HD
authors.lastname: Ellerton
authors.forename: J F
authors.initials: JF
authors.lastname: Koretz
authors.forename: K
authors.initials: K
authors.lastname: Thomson
authors.forename: R C
authors.initials: RC
authors.lastname: Augusteyn
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000378:metabolism*; D005333:Fetus; D007908:Lens, Crystalline / Q000032:analysis; D008854:Microscopy, Electron; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D003459:Crystallins