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Title: The steroidal lactone withaferin A impedes T-cell motility by inhibiting the kinase ZAP70 and subsequent kinome signaling
Authors: Mobashar Hussain Urf Turabe Fazil
Chirumamilla, Chandra Sekhar
Perez-Novo, Claudina
Wong, Brandon Han Siang
Kumar, Sunil
Sze, Siu Kwan
Berghe, Wim Vanden
Verma, Navin Kumar
Keywords: Science::Medicine
Issue Date: 2021
Source: Mobashar Hussain Urf Turabe Fazil, Chirumamilla, C. S., Perez-Novo, C., Wong, B. H. S., Kumar, S., Sze, S. K., Berghe, W. V. & Verma, N. K. (2021). The steroidal lactone withaferin A impedes T-cell motility by inhibiting the kinase ZAP70 and subsequent kinome signaling. Journal of Biological Chemistry, 297(6), 101377-.
Project: MOE2017-T2-2-004
Journal: Journal of Biological Chemistry
Abstract: The steroidal lactone withaferin A (WFA) is a dietary phytochemical, derived from Withania somnifera. It exhibits a wide range of biological properties, including immunomodulatory, anti-inflammatory, antistress, and anticancer activities. Here we investigated the effect of WFA on T-cell motility, which is crucial for adaptive immune responses as well as autoimmune reactions. We found that WFA dose-dependently (within the concentration range of 0.3-1.25 μM) inhibited the ability of human T-cells to migrate via cross-linking of the lymphocyte function-associated antigen-1 (LFA-1) integrin with its ligand, intercellular adhesion molecule 1 (ICAM-1). Coimmunoprecipitation of WFA interacting proteins and subsequent tandem mass spectrometry identified a WFA-interactome consisting of 273 proteins in motile T-cells. In particular, our data revealed significant enrichment of the zeta-chain-associated protein kinase 70 (ZAP70) and cytoskeletal actin protein interaction networks upon stimulation. Phospho-peptide mapping and kinome analysis substantiated kinase signaling downstream of ZAP70 as a key WFA target, which was further confirmed by bait-pulldown and Western immunoblotting assays. The WFA-ZAP70 interaction was disrupted by a disulfide reducing agent dithiothreitol, suggesting an involvement of cysteine covalent binding interface. In silico docking predicted WFA binding to ZAP70 at cystine 560 and 564 residues. These findings provide a mechanistic insight whereby WFA binds to and inhibits the ZAP70 kinase and impedes T-cell motility. We therefore conclude that WFA may be exploited to pharmacologically control host immune responses and potentially prevent autoimmune-mediated pathologies.
ISSN: 0021-9258
DOI: 10.1016/j.jbc.2021.101377
Schools: Lee Kong Chian School of Medicine (LKCMedicine) 
School of Biological Sciences 
Interdisciplinary Graduate School (IGS) 
Research Centres: NTU Institute for Health Technologies 
Rights: © 2021 The Authors. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY license (
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:IGS Journal Articles
LKCMedicine Journal Articles
SBS Journal Articles

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