Direct capture, inhibition and crystal structure of HsaD (Rv3569c) from M. tuberculosis

Autor: Sarah Barelier, Romain Avellan, Giri Raj Gnawali, Patrick Fourquet, Véronique Roig‐Zamboni, Isabelle Poncin, Vanessa Point, Yves Bourne, Stéphane Audebert, Luc Camoin, Christopher D. Spilling, Stéphane Canaan, Jean‐François Cavalier, Gerlind Sulzenbacher
Přispěvatelé: Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire d'ingénierie des systèmes macromoléculaires (LISM), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), University of Missouri [St. Louis], University of Missouri System, Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), ANR-19-CE44-0011,LipInTB,Utilisation de nouveaux inhibiteurs sélectifs pour décrypter le métabolisme lipidique et de la virulence chez M. tb(2019), Cavalier, Jean-François, Utilisation de nouveaux inhibiteurs sélectifs pour décrypter le métabolisme lipidique et de la virulence chez M. tb - - LipInTB2019 - ANR-19-CE44-0011 - AAPG2019 - VALID
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: FEBS Journal
FEBS Journal, 2022, ⟨10.1111/febs.16645⟩
ISSN: 1742-464X
1742-4658
DOI: 10.1111/febs.16645⟩
Popis: International audience; A hallmark of Mycobacterium tuberculosis (M. tb), the etiologic agent of tuberculosis, is its ability to metabolize host-derived lipids. However, the enzymes and mechanisms underlying such metabolism are still largely unknown. We previously reported that the Cyclophostin & Cyclipostins (CyC) analogs, a new family of potent antimycobacterial molecules, react specifically and covalently with (Ser/Cys)-based enzymes mostly involved in bacterial lipid metabolism. Here, we report the synthesis of new CyC alkyne-containing inhibitors (CyCyne) and their use for the direct fishing of target proteins in M. tb culture via bio-orthogonal click-chemistry activity-based protein profiling (CC-ABPP). This approach led to the capture and identification of a variety of enzymes, many of them involved in lipid or steroid metabolisms. One of the captured enzymes, HsaD (Rv3569c), is required for the survival of M. tb within macrophages and is thus a potential therapeutic target. This prompted us to further explore and validate, through a combination of biochemical and structural approaches, the specificity of HsaD inhibition by the CyC analogs. We confirmed that the CyC bind covalently to the catalytic Ser114 residue, leading to a total loss of enzyme activity. These data were supported by the X-ray structures of four HsaD-CyC complexes, obtained at resolutions between 1.6-2.6 Å. The identification of mycobacterial enzymes directly captured by the CyCyne probes through CC-ABPP paves the way to better understand and potentially target key players at crucial stages of the bacilli life-cycle.
Databáze: OpenAIRE
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