Interaction of chaperone alpha-crystallin with unfolded state of alpha-amylase: Implications for reconstitution of the active enzyme.
Alpha-crystallin is reported to act like molecular chaperone by suppressing the aggregation of damaged crystallins in eye lens. In this work, it is shown that alpha-crystallin increases the reactivation of guanidine hydrochloride (GdnHCl)-denatured alpha-amylase from porcine pancreas. 8-Anilinonaphthalene-sulphonate (ANS) binding studies reveal the involvement of hydrophobic interactions in the formation of the complex of alpha-crystallin and alpha-amylase. On the basis of our fluorescence spectroscopic and gel-filtration results, we propose that alpha-crystallin blocks the unfavorable pathways that lead to irreversible denaturation of alpha-amylase and keep it in folding-competent intermediate state.
The Department of Biotechnology and Food Technology, ML Sultan Campus, Durban University of Technology, Durban 4001, Kwa-Zulu Natal, South Africa. ajitkanwal@yahoo.com
Ajit
A
Kumar
Suren
S
Singh
D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D004229:Dithiothreitol / Q000737:chemistry; D019791:Guanidine / Q000737:chemistry; D008027:Light; D010179:Pancreas / Q000378:metabolism*; D011489:Protein Denaturation; D020156:Salicylic Acid / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods; D013379:Substrate Specificity; D013552:Swine; D013997:Time Factors; D000516:alpha-Amylases / Q000737:chemistry*; D038201:alpha-Crystallins / Q000737:chemistry*
D038201:alpha-Crystallins; D000516:alpha-Amylases; D019791:Guanidine; D020156:Salicylic Acid; D004229:Dithiothreitol