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Enzymatic β-Oxidation of the Cholesterol Side Chain in Mycobacterium tuberculosis Bifurcates Stereospecifically at Hydration of 3-Oxo-cholest-4,22-dien-24-oyl-CoA

Journal Article · · ACS Infectious Diseases
 [1];  [2];  [3];  [2];  [4];  [2]
  1. Stony Brook Univ., NY (United States). Dept. of Chemistry; OSTI
  2. Stony Brook Univ., NY (United States). Dept. of Chemistry
  3. Stony Brook Univ., NY (United States). Biochemistry and Structural Biology Graduate Program
  4. Stony Brook Univ., NY (United States). Dept. of Pharmacological Sciences
+ Show Author Affiliations
The unique ability of Mycobacterium tuberculosis (Mtb) to utilize host lipids such as cholesterol for survival, persistence, and virulence has made the metabolic pathway of cholesterol an area of great interest for therapeutics development. Herein, we identify and characterize two genes from the Cho-region (genomic locus responsible for cholesterol catabolism) of the Mtb genome, chsH3 (Rv3538) and chsB1 (Rv3502c). Their protein products catalyze two sequential stereospecific hydration and dehydrogenation steps in the β-oxidation of the cholesterol side chain. ChsH3 favors the 22S hydration of 3-oxo-cholest-4,22-dien24-oyl-CoA in contrast to the previously reported EchA19 (Rv3516), which catalyzes formation of the (22R)-hydroxy-3-oxo-cholest4-en-24-oyl-CoA from the same enoyl-CoA substrate. ChsB1 is stereospecific and catalyzes dehydrogenation of the ChsH3 product but not the EchA19 product. The X-ray crystallographic structure of the ChsB1 apo-protein was determined at a resolution of 2.03 Å, and the holo-enzyme with bound NAD+ cofactor was determined at a resolution of 2.21 Å. The homodimeric structure is representative of a classical NAD+-utilizing short-chain type alcohol dehydrogenase/reductase, including a Rossmann-fold motif, but exhibits a unique substrate binding site architecture that is of greater length and width than its homologous counterparts, likely to accommodate the bulky steroid substrate. Intriguingly, Mtb utilizes hydratases from the MaoC-like family in sterol side-chain catabolism in contrast to fatty acid β-oxidation in other species that utilize the evolutionarily distinct crotonase family of hydratases.
Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division
Grant/Contract Number:
SC0012704
OSTI ID:
1816192
Alternate ID(s):
OSTI ID: 1808965
Journal Information:
ACS Infectious Diseases, Journal Name: ACS Infectious Diseases Journal Issue: 6 Vol. 7; ISSN 2373-8227
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
English

References (47)

The catalytic reaction and inhibition mechanism of Drosophila alcohol dehydrogenase: observation of an enzyme-bound NAD-ketone adduct at 1.4 Å resolution by X-ray crystallography journal June 1999
Sterol carrier protein X is peroxisomal 3-oxoacyl coenzyme A thiolase with intrinsic sterol carrier and lipid transfer activity. journal August 1994
Intermembrane transfer of 5 alpha-cholest-7-en-3 beta-ol. Facilitation by supernatant protein (SCP). journal December 1981
Binary Structure of the Two-Domain (3R)-Hydroxyacyl-CoA Dehydrogenase from Rat Peroxisomal Multifunctional Enzyme Type 2 at 2.38 Å Resolution journal January 2003
More than cholesterol catabolism: regulatory vulnerabilities in Mycobacterium tuberculosis journal June 2018
Metabolomics of Mycobacterium tuberculosis Reveals Compartmentalized Co-Catabolism of Carbon Substrates journal October 2010
Crystal Structure of Yeast Peroxisomal Multifunctional Enzyme: Structural Basis for Substrate Specificity of (3R)-hydroxyacyl-CoA Dehydrogenase Units journal May 2006
FadA5 a Thiolase from Mycobacterium tuberculosis : A Steroid-Binding Pocket Reveals the Potential for Drug Development against Tuberculosis journal January 2015
IpdE1-IpdE2 Is a Heterotetrameric Acyl Coenzyme A Dehydrogenase That Is Widely Distributed in Steroid-Degrading Bacteria journal February 2020
α-Methyl Acyl CoA Racemase Provides Mycobacterium tuberculosis Catabolic Access to Cholesterol Esters journal September 2015
Mycobacterium tuberculosis Exploits a Heterohexameric Enoyl-CoA Hydratase Retro-Aldolase Complex for Cholesterol Catabolism journal September 2019
Hit Generation in TB Drug Discovery: From Genome to Granuloma journal January 2018
Post-translational Succinylation of Mycobacterium tuberculosis Enoyl-CoA Hydratase EchA19 Slows Catalytic Hydration of Cholesterol Catabolite 3-Oxo-chol-4,22-diene-24-oyl-CoA journal July 2020
Mce3R Stress-Resistance Pathway Is Vulnerable to Small-Molecule Targeting That Improves Tuberculosis Drug Activities journal April 2019
Mycobacterium tuberculosis Utilizes a Unique Heterotetrameric Structure for Dehydrogenation of the Cholesterol Side Chain journal April 2013
A Distinct MaoC-like Enoyl-CoA Hydratase Architecture Mediates Cholesterol Catabolism in Mycobacterium tuberculosis journal September 2014
Unraveling Cholesterol Catabolism in Mycobacterium tuberculosis : ChsE4-ChsE5 α 2 β 2 Acyl-CoA Dehydrogenase Initiates β-Oxidation of 3-Oxo-cholest-4-en-26-oyl CoA journal January 2015
Chemical and biological evolution of a nucleotide-binding protein journal July 1974
Unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism journal February 2016
The pursuit of mechanism of action: uncovering drug complexity in TB drug discovery journal January 2021
A gene cluster encoding cholesterol catabolism in a soil actinomycete provides insight into Mycobacterium tuberculosis survival in macrophages journal January 2007
Mycobacterial persistence requires the utilization of host cholesterol journal March 2008
Genetic requirements for mycobacterial survival during infection journal October 2003
Mycobacterial Cytochrome P450 125 (Cyp125) Catalyzes the Terminal Hydroxylation of C27 Steroids journal December 2009
Structural and Biochemical Characterization of Mycobacterium tuberculosis CYP142 journal December 2010
Intracellular Mycobacterium tuberculosis Exploits Host-derived Fatty Acids to Limit Metabolic Stress journal January 2013
Automatic generation of protein structure cartoons with Pro-origami journal October 2011
CoABind: a novel algorithm for Coenzyme A (CoA)- and CoA derivatives-binding residues prediction journal March 2018
Cholesterol and fatty acids grease the wheels of Mycobacterium tuberculosis pathogenesis journal March 2018
Role of the methylcitrate cycle in propionate metabolism and detoxification in Mycobacterium smegmatis journal December 2007
XDS journal January 2010
Data processing and analysis with the autoPROC toolbox journal March 2011
How good are my data and what is the resolution? journal June 2013
A patchwork pathway for oxygenase-independent degradation of side chain containing steroids: Anaerobic degradation of steroids journal November 2017
Steryl Ester Formation and Accumulation in Steroid-Degrading Bacteria journal November 2019
Functional Characterization of Three Specific Acyl-Coenzyme A Synthetases Involved in Anaerobic Cholesterol Degradation in Sterolibacterium denitrificans Chol1S journal April 2018
A Thiolase of Mycobacterium tuberculosis Is Required for Virulence and Production of Androstenedione and Androstadienedione from Cholesterol journal January 2010
The Essential Function of Genes for a Hydratase and an Aldehyde Dehydrogenase for Growth of Pseudomonas sp. Strain Chol1 with the Steroid Compound Cholate Indicates an Aldolytic Reaction Step for Deacetylation of the Side Chain journal August 2013
Shrinking the FadE Proteome of Mycobacterium tuberculosis: Insights into Cholesterol Metabolism through Identification of an α 2 β 2 Heterotetrameric Acyl Coenzyme A Dehydrogenase Family journal October 2013
Characterization of an Aldolase Involved in Cholesterol Side Chain Degradation in Mycobacterium tuberculosis journal January 2018
Actinobacterial Acyl Coenzyme A Synthetases Involved in Steroid Side-Chain Catabolism journal February 2014
Functional Characterization of a Vitamin B12-Dependent Methylmalonyl Pathway in Mycobacterium tuberculosis: Implications for Propionate Metabolism during Growth on Fatty Acids journal March 2008
Identification and Characterization of a New Enoyl Coenzyme A Hydratase Involved in Biosynthesis of Medium-Chain-Length Polyhydroxyalkanoates in Recombinant Escherichia coli journal September 2003
Integration of Metabolomics and Transcriptomics Reveals a Complex Diet of Mycobacterium tuberculosis during Early Macrophage Infection journal August 2017
High-Resolution Phenotypic Profiling Defines Genes Essential for Mycobacterial Growth and Cholesterol Catabolism journal September 2011
Novel Inhibitors of Cholesterol Degradation in Mycobacterium tuberculosis Reveal How the Bacterium’s Metabolism Is Constrained by the Intracellular Environment journal February 2015
Pathogen roid rage: Cholesterol utilization by Mycobacterium tuberculosis journal March 2014

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Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
Mycobacterium tuberculosis
Rv3502c
Rv3538
catabolism
cholesterol
conformation
degradation
hydration
peptides and proteins
stereochemistry