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Alpha-Glucosidase Folding During Urea Denaturation: Enzyme Kinetics and Computational Prediction

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Abstract

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.

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Abbreviations

pNPG:

p-Nitrophenyl α-d-glucopyranoside

pNP:

4-Nitrophenol

SDS:

Sodium dodecyl sulfate

ANS:

1-Anilino-8-naphthalenesulfonate

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Acknowledgements

Dr. Fei Zou was supported by a grant from the National Basic Research Program of China (no. 2006CB504100). Dr. Jong Bhak was supported by a grant from the KRIBB Research Initiative Program of Korea. Dr. Yong-Doo Park was supported by fund from the Science and Technology Planning Project of Jiaxing (no. 2008AZ1024), Zhejiang.

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Authors and Affiliations

  1. 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

    Xue-Qiang Wu, Zhi-Rong Lü, Long Shi, Yong-Doo Park & Fei Zou

  2. School of Medicine, Shenzhen University, Shenzhen, 518060, People’s Republic of China

    Jun Wang

  3. Yangtze Delta Region Institute of Tsinghua University, Jiaxing, 314050, People’s Republic of China

    Hong-Min Tang & Yong-Doo Park

  4. Korean BioInformation Center (KOBIC), KRIBB, Daejeon, 305-806, South Korea

    Daeui Park, Sang-Ho Oh & Jong Bhak

Authors
  1. Xue-Qiang Wu
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  2. Jun Wang
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  3. Zhi-Rong Lü
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  4. Hong-Min Tang
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  7. Jong Bhak
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  8. Long Shi
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  9. Yong-Doo Park
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  10. Fei Zou
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Correspondence to Fei Zou.

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Xue-Qiang Wu and Jun Wang have contributed equally to this study.

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Wu, XQ., Wang, J., Lü, ZR. et al. Alpha-Glucosidase Folding During Urea Denaturation: Enzyme Kinetics and Computational Prediction. Appl Biochem Biotechnol 160, 1341–1355 (2010). https://doi.org/10.1007/s12010-009-8636-6

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