title: Effects of macromolecular crowding on the refolding of glucose- 6-phosphate dehydrogenase and protein disulfide isomerase.
abstract: The effects of polysaccharide, polyethylene glycol, and protein-crowding agents on the refolding of glucose-6-phosphate dehydrogenase (G6PDH) and protein disulfide isomerase have been examined. By increasing concentration during refolding, the reactivation yields of the two proteins decrease with the formation of soluble aggregates. In the presence of high concentrations of crowding agents the reactivation yields remain constant but with decreased refolding rates. The refolding of G6PDH changes from monophasic to biphasic first-order reactions in the presence of crowding agents, and the amplitude of the new slow phase increases with increasing concentrations of crowding agents. The molecular chaperone GroEL reverses the refolding kinetics of G6PDH from biphase back to monophase and accelerates the refolding process. Our results display the complexity and diversity of the effects of macromolecular crowding on both the thermodynamics and kinetics of protein folding.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, 15 Datun Road, Beijing 100101, China.
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: S
authors.initials: S
authors.lastname: Zhang
authors.forename: C
authors.initials: C
authors.lastname: Wang
mesh_terms: D018834:Chaperonin 60 / Q000502:physiology; D004789:Enzyme Activation; D005954:Glucosephosphate Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D009113:Muramidase / Q000494:pharmacology; D010047:Ovalbumin / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D019704:Protein Disulfide-Isomerases / Q000737:chemistry* / Q000378:metabolism; D017510:Protein Folding; D012710:Serum Albumin, Bovine / Q000494:pharmacology
chemical_list: D018834:Chaperonin 60; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; D010047:Ovalbumin; D005954:Glucosephosphate Dehydrogenase; D009113:Muramidase; D019704:Protein Disulfide-Isomerases