High-resolution crystal structure of human asparagine synthetase enables analysis of inhibitor binding and selectivity

Abstract

Expression of human asparagine synthetase (ASNS) promotes metastatic progression and tumor cell invasiveness in colorectal and breast cancer, presumably by altering cellular levels of L-asparagine. Human ASNS is therefore emerging as a bona fide drug target for cancer therapy. Here we show that a slow-onset, tight binding inhibitor, which exhibits nanomolar affinity for human ASNS in vitro, exhibits excellent selectivity at 10 μM concentration in HCT-116 cell lysates with almost no off-target binding. The high-resolution (1.85 Å) crystal structure of human ASNS has enabled us to identify a cluster of negatively charged side chains in the synthetase domain that plays a key role in inhibitor binding. Comparing this structure with those of evolutionarily related AMP-forming enzymes provides insights into intermolecular interactions that give rise to the observed binding selectivity. Our findings demonstrate the feasibility of developing second generation human ASNS inhibitors as lead compounds for the discovery of drugs against metastasis.

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Item Type:	Article
Authors/Creators:	Zhu, W. Radadiya, A. Bisson, C. https://orcid.org/0000-0002-9430-6822 Wenzel, S. Nordin, B.E. Martínez-Márquez, F. Imasaki, T. Sedelnikova, S.E. https://orcid.org/0000-0001-5174-6302 Coricello, A. Baumann, P. Berry, A.H. Nomanbhoy, T.K. Kozarich, J.W. Jin, Y. Rice, D.W. https://orcid.org/0000-0002-7811-0539 Takagi, Y. Richards, N.G.J.
Copyright, Publisher and Additional Information:	© 2019 The Authors. This article is licensed under a Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/.), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
Keywords:	Computational models; Enzymes; Medicinal chemistry; X-ray crystallography
Dates:	Accepted: 21 August 2019 Published: 17 September 2019
Institution:	The University of Sheffield
Academic Units:	The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) > Department of Molecular Biology and Biotechnology (Sheffield)
Depositing User:	Symplectic Sheffield
Date Deposited:	30 Oct 2019 13:32
Last Modified:	18 Mar 2020 15:41
Status:	Published
Publisher:	Springer Science and Business Media LLC
Refereed:	Yes
Identification Number:	10.1038/s42003-019-0587-z
Related URLs:	Erratum/Corrigendum
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:152720

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High-resolution crystal structure of human asparagine synthetase enables analysis of inhibitor binding and selectivity

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