A non-traditional crystal-based compound screening method targeting the ATP binding site of GRP78 for identification of novel nucleoside analogues

Authors

Alexander Mrozek, Department of Biochemistry & Molecular Biology, Tulane University School of Medicine, New Orleans, LA, United States.
Tetyana Antoshchenko, Department of Biochemistry & Molecular Biology, Tulane University School of Medicine, New Orleans, LA, United States.
Yun Chen, Department of Biochemistry & Molecular Biology, Tulane University School of Medicine, New Orleans, LA, United States.
Carlos Zepeda-Velázquez, Drug Discovery Program, Ontario Institute for Cancer Research, Toronto, ON, Canada.
David Smil, Drug Discovery Program, Ontario Institute for Cancer Research, Toronto, ON, Canada.
Nirbhay Kumar, Department of Global Health, George Washington University Milken Institute of Public Health, Washington, D.C., DC, United States.
Hua Lu, Department of Biochemistry & Molecular Biology, Tulane University School of Medicine, New Orleans, LA, United States.
Hee-Won Park, Department of Biochemistry & Molecular Biology, Tulane University School of Medicine, New Orleans, LA, United States.

Document Type

Journal Article

Publication Date

1-1-2022

Journal

Frontiers in molecular biosciences

Volume

9

DOI

10.3389/fmolb.2022.956095

Keywords

GRP78; Plasmodium falciparum; chaperone; drug discovery & development; malaria; nucleoside analogues; unfolded protein response

Abstract

Drug resistance to front-line malarial treatments represents an ongoing threat to control malaria, a vector borne infectious disease. The malarial parasite, has developed genetic variants, conferring resistance to the current standard therapeutic artemisinin and its derivatives commonly referred to as artemisinin-combination therapies (ACTs). Emergence of multi-drug resistance parasite genotypes is a warning of potential treatment failure, reaffirming the urgent and critical need to find and validate alternate drug targets to prevent the spread of disease. An attractive and novel drug target includes glucose-regulated protein 78 kDa (GRP78, or BiP), an essential molecular chaperone protein involved in the unfolded protein response that is upregulated in ACT treated parasites. We have shown that both sequence and structure are closely related to human GRP78 (hGRP78), a chaperone belonging to the HSP70 class of ATPase proteins, which is often upregulated in cellular stress responses and cancer. By screening a library of nucleoside analogues, we identified eight 'hit' compounds binding at the active site of the ATP binding domain of GRP78 using a high-throughput ligand soaking screen using x-ray crystallography. These compounds were further evaluated using protein thermal shift assays to assess target binding activity. The nucleoside analogues identified from our screen provide a starting point for the development of more potent and selective antimalarial inhibitors. In addition, we have established a well-defined, high-throughput crystal-based screening approach that can be applied to many crystallizable proteins for generating anti- specific compounds.

APA Citation

Mrozek, Alexander; Antoshchenko, Tetyana; Chen, Yun; Zepeda-Velázquez, Carlos; Smil, David; Kumar, Nirbhay; Lu, Hua; and Park, Hee-Won, "A non-traditional crystal-based compound screening method targeting the ATP binding site of GRP78 for identification of novel nucleoside analogues" (2022). GW Authored Works. Paper 1805.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/1805

Department

Global Health