Mycobacterium tuberculosis Rv1916 is an Acetyl-CoA-Binding Protein

Evelyn Yu Wen Huang; Brooke X.C. Kwai; Ram Prasad Bhusal; Ghader Bashiri; Ivanhoe K.H. Leung

doi:10.1002/cbic.202300162

Mycobacterium tuberculosis Rv1916 is an Acetyl-CoA-Binding Protein

Evelyn Yu Wen Huang, Brooke X.C. Kwai, Ram Prasad Bhusal, Ghader Bashiri, Ivanhoe K.H. Leung

Research output: Contribution to journal › Article › Research › peer-review

2 Citations (Scopus)

Abstract

Isocitrate lyase (ICL) isoform 2 is an essential enzyme for some clinical Mycobacterium tuberculosis (Mtb) strains during infection. In the laboratory Mtb strain H37Rv, the icl2 gene encodes two distinct gene products – Rv1915 and Rv1916 – due to a frameshift mutation. This study aims to characterise these two gene products to understand their structure and function. While we were unable to produce Rv1915 recombinantly, soluble Rv1916 was obtained with sufficient yield for characterisation. Kinetic studies using UV-visible spectrophotometry and ¹H-NMR spectroscopy showed that recombinant Rv1916 does not possess isocitrate lyase activity, while waterLOGSY binding experiments demonstrated that it could bind acetyl-CoA. Finally, X-ray crystallography revealed structural similarities between Rv1916 and the C-terminal domain of ICL2. Considering the probable differences between full-length ICL2 and the gene products Rv1915 and Rv1916, care must be taken when using Mtb H37Rv as a model organism to study central carbon metabolism.

Original language	English
Article number	e202300162
Number of pages	7
Journal	ChemBioChem
Volume	24
Issue number	14
DOIs	https://doi.org/10.1002/cbic.202300162
Publication status	Published - 13 Jul 2023

Keywords

acyl-CoA-binding protein
glyoxylate shunt
isocitrate lyase
Mycobacterium tuberculosis
Rv1916

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/cbic.202300162Licence: CC BY-NC-ND

Cite this

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title = "Mycobacterium tuberculosis Rv1916 is an Acetyl-CoA-Binding Protein",

abstract = "Isocitrate lyase (ICL) isoform 2 is an essential enzyme for some clinical Mycobacterium tuberculosis (Mtb) strains during infection. In the laboratory Mtb strain H37Rv, the icl2 gene encodes two distinct gene products – Rv1915 and Rv1916 – due to a frameshift mutation. This study aims to characterise these two gene products to understand their structure and function. While we were unable to produce Rv1915 recombinantly, soluble Rv1916 was obtained with sufficient yield for characterisation. Kinetic studies using UV-visible spectrophotometry and 1H-NMR spectroscopy showed that recombinant Rv1916 does not possess isocitrate lyase activity, while waterLOGSY binding experiments demonstrated that it could bind acetyl-CoA. Finally, X-ray crystallography revealed structural similarities between Rv1916 and the C-terminal domain of ICL2. Considering the probable differences between full-length ICL2 and the gene products Rv1915 and Rv1916, care must be taken when using Mtb H37Rv as a model organism to study central carbon metabolism.",

keywords = "acyl-CoA-binding protein, glyoxylate shunt, isocitrate lyase, Mycobacterium tuberculosis, Rv1916",

author = "Huang, {Evelyn Yu Wen} and Kwai, {Brooke X.C.} and Bhusal, {Ram Prasad} and Ghader Bashiri and Leung, {Ivanhoe K.H.}",

note = "Funding Information: EYH acknowledges the financial support received from the University of Melbourne through the Melbourne Research Scholarship and Rowden White Scholarship. RPB acknowledges the financial support received from the University of Auckland through the University of Auckland Doctoral Scholarship. GB and IKHL acknowledge the financial support received from the University of Auckland. GB is supported by a Sir Charles Hercus Fellowship through the Health Research Council of New Zealand. BXCK and IKHL acknowledge the financial support received from the University of Melbourne through the Driving Research Momentum initiative. This research was undertaken on the MX2 beamline at the Australian Synchrotron, part of ANSTO. Open Access publishing facilitated by The University of Melbourne, as part of the Wiley ‐ The University of Melbourne agreement via the Council of Australian University Librarians. [35] Publisher Copyright: {\textcopyright} 2023 The Authors. ChemBioChem published by Wiley-VCH GmbH.",

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AU - Huang, Evelyn Yu Wen

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AU - Bhusal, Ram Prasad

AU - Bashiri, Ghader

AU - Leung, Ivanhoe K.H.

N1 - Funding Information: EYH acknowledges the financial support received from the University of Melbourne through the Melbourne Research Scholarship and Rowden White Scholarship. RPB acknowledges the financial support received from the University of Auckland through the University of Auckland Doctoral Scholarship. GB and IKHL acknowledge the financial support received from the University of Auckland. GB is supported by a Sir Charles Hercus Fellowship through the Health Research Council of New Zealand. BXCK and IKHL acknowledge the financial support received from the University of Melbourne through the Driving Research Momentum initiative. This research was undertaken on the MX2 beamline at the Australian Synchrotron, part of ANSTO. Open Access publishing facilitated by The University of Melbourne, as part of the Wiley ‐ The University of Melbourne agreement via the Council of Australian University Librarians. [35] Publisher Copyright: © 2023 The Authors. ChemBioChem published by Wiley-VCH GmbH.

PY - 2023/7/13

Y1 - 2023/7/13

N2 - Isocitrate lyase (ICL) isoform 2 is an essential enzyme for some clinical Mycobacterium tuberculosis (Mtb) strains during infection. In the laboratory Mtb strain H37Rv, the icl2 gene encodes two distinct gene products – Rv1915 and Rv1916 – due to a frameshift mutation. This study aims to characterise these two gene products to understand their structure and function. While we were unable to produce Rv1915 recombinantly, soluble Rv1916 was obtained with sufficient yield for characterisation. Kinetic studies using UV-visible spectrophotometry and 1H-NMR spectroscopy showed that recombinant Rv1916 does not possess isocitrate lyase activity, while waterLOGSY binding experiments demonstrated that it could bind acetyl-CoA. Finally, X-ray crystallography revealed structural similarities between Rv1916 and the C-terminal domain of ICL2. Considering the probable differences between full-length ICL2 and the gene products Rv1915 and Rv1916, care must be taken when using Mtb H37Rv as a model organism to study central carbon metabolism.

AB - Isocitrate lyase (ICL) isoform 2 is an essential enzyme for some clinical Mycobacterium tuberculosis (Mtb) strains during infection. In the laboratory Mtb strain H37Rv, the icl2 gene encodes two distinct gene products – Rv1915 and Rv1916 – due to a frameshift mutation. This study aims to characterise these two gene products to understand their structure and function. While we were unable to produce Rv1915 recombinantly, soluble Rv1916 was obtained with sufficient yield for characterisation. Kinetic studies using UV-visible spectrophotometry and 1H-NMR spectroscopy showed that recombinant Rv1916 does not possess isocitrate lyase activity, while waterLOGSY binding experiments demonstrated that it could bind acetyl-CoA. Finally, X-ray crystallography revealed structural similarities between Rv1916 and the C-terminal domain of ICL2. Considering the probable differences between full-length ICL2 and the gene products Rv1915 and Rv1916, care must be taken when using Mtb H37Rv as a model organism to study central carbon metabolism.

KW - acyl-CoA-binding protein

KW - glyoxylate shunt

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KW - Mycobacterium tuberculosis

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DO - 10.1002/cbic.202300162

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JO - ChemBioChem

JF - ChemBioChem

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M1 - e202300162

ER -

Mycobacterium tuberculosis Rv1916 is an Acetyl-CoA-Binding Protein

Abstract

Keywords

UN SDGs

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Equipment

Australian Synchrotron

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