Alpha-glucosidase folding during urea denaturation: enzyme kinetics and computational prediction.
In this study, we investigated structural changes in alpha-glucosidase during urea denaturation. Alpha-glucosidase was inactivated by urea in a dose-dependent manner. The inactivation was a first-order reaction with a monophase process. Urea inhibited alpha-glucosidase in a mixed-type reaction. We found that an increase in the hydrophobic surface of this enzyme induced by urea resulted in aggregation caused by unstable folding intermediates. We also simulated the docking between alpha-glucosidase and urea. The docking simulation suggested that several residues, namely THR9, TRP14, LYS15, THR287, ALA289, ASP338, SER339, and TRP340, interact with urea. Our study provides insights into the alpha-glucosidase unfolding pathway and 3D structure of alpha-glucosidase.
Wu
Xue-Qiang
XQ
Department of Environmental Health, School of Public Health and Tropical Medicine, Southern Medical University, Baiyun District, North of Guangzhou Road No.1838, Guangzhou 510515, People's Republic of China.
Wang
Jun
J
Lü
Zhi-Rong
ZR
Tang
Hong-Min
HM
Park
Daeui
D
Oh
Sang-Ho
SH
Bhak
Jong
J
Shi
Long
L
Park
Yong-Doo
YD
Zou
Fei
F
D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D019295:Computational Biology; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D056004:Molecular Dynamics Simulation; D011489:Protein Denaturation / Q000187:drug effects*; D017510:Protein Folding / Q000187:drug effects*; D020836:Protein Structure, Quaternary; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014508:Urea / Q000494:pharmacology*; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000378:metabolism*
D000817:Anilino Naphthalenesulfonates; D012996:Solutions; C027132:1-anilino-8-naphthalenesulfonate; D014508:Urea; D000520:alpha-Glucosidases