University of Oulu

Dalwani, S., Lampela, O., Leprovost, P., Schmitz, W., Juffer, A. H., Wierenga, R. K., & Venkatesan, R. (2021). Substrate specificity and conformational flexibility properties of the Mycobacterium tuberculosis β-oxidation trifunctional enzyme. Journal of Structural Biology, 213(3), 107776.

Substrate specificity and conformational flexibility properties of the Mycobacterium tuberculosis β-oxidation trifunctional enzyme

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Author: Dalwani, Subhadra1; Lampela, Outi1,2; Leprovost, Pierre1,2;
Organizations: 1Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
2Biocenter Oulu, University of Oulu, Oulu, Finland
3Theoder-Boveri-Institut für Biowissenschaften der Universität Würzburg, Würzburg, Germany
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 9.3 MB)
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Language: English
Published: Elsevier, 2021
Publish Date: 2021-09-15


The Mycobacterium tuberculosis trifunctional enzyme (MtTFE) is an α₂β₂ tetrameric enzyme. The α-chain harbors the 2E-enoyl-CoA hydratase (ECH) and 3S-hydroxyacyl-CoA dehydrogenase (HAD) activities and the β-chain provides the 3-ketoacyl-CoA thiolase (KAT) activity. Enzyme kinetic data reported here show that medium and long chain enoyl-CoA molecules are preferred substrates for MtTFE. Modelling studies indicate how the linear medium and long acyl chains of these substrates can bind to each of the active sites. In addition, crystallographic binding studies have identified three new CoA binding sites which are different from the previously known CoA binding sites of the three TFE active sites. Structure comparisons provide new insights into the properties of ECH, HAD and KAT active sites of MtTFE. The interactions of the adenine moiety of CoA with loop-2 of the ECH active site cause a conformational change of this loop by which a competent ECH active site is formed. The NAD+ binding domain (domain C) of the HAD part of MtTFE has only a few interactions with the rest of the complex and adopts a range of open conformations, whereas the A-domain of the ECH part is rigidly fixed with respect to the HAD part. Two loops, the CB1-CA1 region and the catalytic CB4-CB5 loop, near the thiolase active site and the thiolase dimer interface, have high B-factors. Structure comparisons suggest that a competent and stable thiolase dimer is formed only when complexed with the α-chains, highlighting the importance of the assembly for the proper functioning of the complex.

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Series: Journal of structural biology
ISSN: 1047-8477
ISSN-E: 1095-8657
ISSN-L: 1047-8477
Volume: 213
Issue: 3
Article number: 107776
DOI: 10.1016/j.jsb.2021.107776
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Funding: This research has been supported by grants of the Academy of Finland (293369, 289024 and 319194) to RV and SD.
Academy of Finland Grant Number: 293369
Detailed Information: 293369 (Academy of Finland Funding decision)
289024 (Academy of Finland Funding decision)
319194 (Academy of Finland Funding decision)
Dataset Reference: Supplementary data:
Copyright information: © 2021 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (