Stimulation of GMP formation in hGBP1 is mediated by W79 and its effect on the antiviral activity

Raninga, N and Nayeem, SM and Gupta, S and Mullick, R and Pandita, E and Das, S and Deep, S and Sau, AK (2020) Stimulation of GMP formation in hGBP1 is mediated by W79 and its effect on the antiviral activity. In: FEBS Journal . (In Press)

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Abstract

Interferon-inducible large GTPases are critical for innate immunity. The distinctive feature of a large GTPase, human guanylate binding protein-1 (hGBP1), is the sequential hydrolysis of GTP into GMP via GDP. Despite several structural and biochemical studies, the underlying mechanism of assembly-stimulated GMP formation by hGBP1 and its role in immunity are not fully clarified. Using a series of biochemical, biophysical, and in silico experiments, we studied four tryptophan residues, located near switch I-II (in and around the active site) to understand the conformational changes near these regions and also to investigate their effect on enhanced GMP formation. The W79A mutation showed significantly reduced GMP formation, whereas the W81A and W180A substitutions exhibited only a marginal defect. The W114A mutation showed a long-range effect of further enhanced GMP formation, which was mediated through W79. We also observed that after first phosphate cleavage, the W79-containing region undergoes a conformational change, which is essential for stimulated GMP formation. We suggest that this conformational change helps to reposition the active site for the next cleavage step, which occurs through a stable contact between the indole moiety of W79 and the main chain carbonyl of K76. We also showed that stimulated GMP formation is crucial for antiviral activity against hepatitis C. Thus, the present study not only provides new insight for the stimulation of GMP formation in hGBP1, but also highlights the importance of the enhanced second phosphate cleavage product in the antiviral activity. © 2020 Federation of European Biochemical Societies

Item Type:	Journal Article
Publication:	FEBS Journal
Publisher:	Blackwell Publishing Ltd
Additional Information:	Copyright to this article belongs to Blackwell Publishing Ltd
Keywords:	conformational change; fluorescencedecays; large GTPase; mutational studies
Department/Centre:	Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited:	11 Dec 2020 09:42
Last Modified:	11 Dec 2020 09:42
URI:	http://eprints.iisc.ac.in/id/eprint/67237

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