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Insights into pH-Induced Conformational Transition of β-Galactosidase from Pisum sativum Leading to its Multimerization

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Abstract

Although β-galactosidases are physiologically a very important enzyme and have may therapeutics applications, very little is known about the stability and the folding aspects of the enzyme. We have used β-galactosidase from Pisum sativum (PsBGAL) as model system to investigate stability, folding, and function relationship of β-galactosidases. PsBGAL is a vacuolar protein which has a tendency to multimerize at acidic pH with protein concentration ≥100 μg mL−1 and dissociates into its subunits above neutral pH. It exhibits maximum activity as well as stability under acidic conditions. Further, it has different conformational orientations and core secondary structures at different pH. Substantial predominance of β-content and interfacial interactions through Trp residues play crucial role in pH-dependent multimerization of enzyme. Equilibrium unfolding of PsBGAL at acidic pH follows four-state model when monitored by changes in the secondary structure with two intermediates: one resembling to molten globule-like state while unfolding seen from activity and tertiary structure of PsBGAL fits to two-state model. Unfolding of PsBGAL at higher pH always follows two-state model. Furthermore, unfolding of PsBGAL reveals that it has at least two domains: α/β barrel containing catalytic site and the other is rich in β-content responsible for enzyme multimerization.

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Acknowledgement

A.D. would like to thank the Council of Scientific and Industrial Research (CSIR), New Delhi for financial assistance in the form of research fellowship.

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Authors and Affiliations

  1. School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi, 221005, India

    Alka Dwevedi & Arvind M. Kayastha

  2. Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Vikash Kumar Dubey

  3. Molecular Biology Unit, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India

    Medicherla V. Jagannadham

Authors
  1. Alka Dwevedi
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  2. Vikash Kumar Dubey
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  3. Medicherla V. Jagannadham
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  4. Arvind M. Kayastha
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Correspondence to Arvind M. Kayastha.

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Dwevedi, A., Dubey, V.K., Jagannadham, M.V. et al. Insights into pH-Induced Conformational Transition of β-Galactosidase from Pisum sativum Leading to its Multimerization. Appl Biochem Biotechnol 162, 2294–2312 (2010). https://doi.org/10.1007/s12010-010-9003-3

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  • DOI: https://doi.org/10.1007/s12010-010-9003-3

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