Structural elucidation of the binding site and mode of inhibition of Li+ and Mg2+ in inositol monophosphatase

Anirudha Dutta; Sudipta Bhattacharyya; Debajyoti Dutta; Amit Kumar Das

doi:10.1111/febs.13070

journal article Oct 27, 2014

Structural elucidation of the binding site and mode of inhibition of Li+ and Mg2+ in inositol monophosphatase

Anirudha Dutta Sudipta Bhattacharyya Debajyoti Dutta Amit Kumar Das

The FEBS Journal Vol. 281 No. 23 pp. 5309-5324 · Wiley

View at Publisher Save 10.1111/febs.13070

Abstract

Mg2+‐dependent, Li+‐sensitive phosphatases are a widely distributed family of enzymes with significant importance throughout the biological kingdom. Inositol monophosphatase (IMPase) is an important target of Li+‐based therapeutic agents in manic depressive disorders. However, despite decades of intense research efforts, the precise mechanism of Li+‐induced inhibition of IMPase remains obscured. Here we describe a structural investigation of the Li+ binding site in staphylococcal IMPase I (SaIMPase I) using X–ray crystallography. The biochemical study indicated common or overlapping binding sites for Mg2+ and Li+ in the active site of SaIMPase I. The crystal structure of the SaIMPase I ternary product complex shows the presence of a phosphate and three Mg2+ ions (namely Mg1, Mg2 and Mg3) in the active site. As Li+ is virtually invisible in X–ray crystallography, competitive displacement of Mg2+ ions from the SaIMPase I ternary product complex as a function of increasing LiCl concentration was used to identify the Li+ binding site. In this approach, the disappearing electron density of Mg2+ ions due to Li+ ion binding was traced, and the Mg2+ ion present at the Mg2 binding site was found to be replaced. Moreover, based on a detailed comparative investigation of the phosphate orientation and coordination states of Mg2+ binding sites in enzyme–substrate and enzyme–product complexes, inhibition mechanisms for Li+ and Mg2+ are proposed.DatabaseThe atomic coordinates for the SaIMPase I ternary complex, SaIMPase I in 50 mm LiCl, SaIMPase I in 100 mm LiCl and SaIMPase I in 0 mm MgCl2 have been submitted to the Protein Data Bank under accession numbers 4G61, 4I40, 4I3Y and 4PTK, respectively.Structured digital abstract

SaIMPase-I and SaIMPase-I bind by x-ray crystallography (View interaction)

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Details

Published: Oct 27, 2014
Vol/Issue: 281(23)
Pages: 5309-5324
License: View

Authors

A

Anirudha Dutta

Department of Biotechnology Indian Institute of Technology Kharagpur Kharagpur West Bengal India

S

Sudipta Bhattacharyya

Department of Biotechnology Indian Institute of Technology Kharagpur Kharagpur West Bengal India

D

Debajyoti Dutta

Department of Biotechnology Indian Institute of Technology Kharagpur Kharagpur West Bengal India

A

Amit Kumar Das

Department of Biotechnology Indian Institute of Technology Kharagpur Kharagpur West Bengal India

Funding

Department of Science and Technology (DST), Government of India Award: SR/SO/BB-065/2008

Cite This Article

Anirudha Dutta, Sudipta Bhattacharyya, Debajyoti Dutta, et al. (2014). Structural elucidation of the binding site and mode of inhibition of Li+ and Mg2+ in inositol monophosphatase. The FEBS Journal, 281(23), 5309-5324. https://doi.org/10.1111/febs.13070

Structural elucidation of the binding site and mode of inhibition of Li+ and Mg2+ in inositol monophosphatase

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