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Tailoring the pH Dependence of Human Non-pancreatic Secretory Phospholipase A2 by Engineering Surface Charges

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Abstract

Human non-pancreatic secretory phospholipase A2 (hnpsPLA2) catalyzes the sn-2 acyl hydrolysis of phospholipids. It was reported that hnpsPLA2 is involved in various diseases like inflammation, cancer, and so on. This enzyme also exhibits anti-bacterial and anti-virus activities. It is active over a broad pH range, with higher activity at alkaline conditions. In order to make it suitable as a potential bactericide, high activity at neutral pH is preferable. We have tried to tailor the pH dependence of hnpsPLA2 activity by replacing its surface charged residues. Three surface charge replacements, Arg42Glu, Arg100Glu, and Glu89Lys, showed increased activities at neutral pH, which are 2.3, 2.8, and 2.3 times that of the wild-type enzyme at pH 7. Both the positive-to-negative and negative-to-positive mutations lowered the optimum enzymatic reaction pH of hnpsPLA2, indicating that the enzyme pH profile depends on a delicate balance of charged residues. The activity changes are in good agreement with the recently proposed calcium-coordinated catalytic triad mechanism. This study also provides a general means of enhancing hnpsPLA2 activity at low pH.

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Abbreviations

hnpsPLA2 :

Human non-pancreatic secretory phospholipase A2

IBS:

Interface binding site

CD:

Circular dichroism

ANS:

1-anilinonaphthalene-8-sulfonate

DMPC:

1,2-dimyristoyl-sn-glycero-3-phosphocholine

GIIAPLA2 :

Group IIA phospholipase A2

V max :

The maximum velocity of the enzymatic activity

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Acknowledgments

This work was supported in part by the National Natural Science Foundation of China and the Ministry of Science and Technology of China.

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Authors and Affiliations

  1. BNLMS, State Key Laboratory of Structural Chemistry for Unstable and Stable Species, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China

    Shunchen Qiu & Luhua Lai

  2. Center for Quantitative Biology, Peking University, Beijing, 100871, China

    Luhua Lai

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  1. Shunchen Qiu
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  2. Luhua Lai
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Correspondence to Luhua Lai.

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Qiu, S., Lai, L. Tailoring the pH Dependence of Human Non-pancreatic Secretory Phospholipase A2 by Engineering Surface Charges. Appl Biochem Biotechnol 171, 1454–1464 (2013). https://doi.org/10.1007/s12010-013-0437-2

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  • DOI: https://doi.org/10.1007/s12010-013-0437-2

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