Comparative studies of the artificial chaperone-assisted refolding of thermally denatured bovine carbonic anhydrase using different capturing ionic detergents and beta-cyclodextrin.
Artificial chaperone-assisted refolding has been shown to be an effective approach for improving the refolding yield of some of the denatured proteins. Since identical concentrations of various detergents do not induce similar variations in the protein structures, we arranged to evaluate the artificial chaperoning capabilities of several ionic detergents as a function of charge, structure, and the hydrophobic tail length of the detergent. Our results indicate that carbonic anhydrase can be refolded from its denatured state via artificial chaperone strategy using both anionic and cationic detergents. However, the extent of refolding assistance (kinetic and refolding yield) were different due to protein and detergent net charges, detergent concentrations, and the length of hydrophobic portion of each detergent. These observed differences were attributed to physical properties of CA-detergent complexes and/or to the kinetics of detergent stripping by beta-cyclodextrin from the protein-detergent complexes which is apparently dependent on the detergent-beta-CD association constants and the nature of the partially stripped complexes.
Yazdanparast
Razieh
R
Institute of Biochemistry and Biophysics, University of Tehran, P.O. Box 13145-1384, Tehran, Iran. yazdan@ibb.ut.ac.ir
Khodarahmi
Reza
R
Soori
Effat
E
D000818:Animals; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D002942:Circular Dichroism; D003902:Detergents / Q000737:chemistry / Q000494:pharmacology*; D007700:Kinetics; D018832:Molecular Chaperones / Q000737:chemistry / Q000494:pharmacology*; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation / Q000187:drug effects; D013050:Spectrometry, Fluorescence; D013696:Temperature; D047392:beta-Cyclodextrins / Q000494:pharmacology*
D003902:Detergents; D018832:Molecular Chaperones; D047392:beta-Cyclodextrins; D002256:Carbonic Anhydrases