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

Chembiochem. 2023 Jul 17;24(14):e202300162. doi: 10.1002/cbic.202300162. Epub 2023 Jun 18.

Authors

Evelyn Yu-Wen Huang¹, Brooke X C Kwai^{1

2

3}, Ram Prasad Bhusal^{2

4}, Ghader Bashiri⁵, Ivanhoe K H Leung^{1

2}

Affiliations

¹ School of Chemistry and the Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, 30 Flemington Rd, Parkville, VIC 3052, Australia.
² School of Chemical Sciences, The University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand.
³ Present address: Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC 3052, Australia.
⁴ Present address: Monash Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800, Australia.
⁵ School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand.

PMID: 37211532
DOI: 10.1002/cbic.202300162

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 ¹ H-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: Mycobacterium tuberculosis; Rv1916; acyl-CoA-binding protein; glyoxylate shunt; isocitrate lyase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acetyl Coenzyme A
Bacterial Proteins / metabolism
Isocitrate Lyase / chemistry
Isocitrate Lyase / genetics
Isocitrate Lyase / metabolism
Kinetics
Mycobacterium tuberculosis*

Substances

Acetyl Coenzyme A
Isocitrate Lyase
Bacterial Proteins