Expression and characterization of Mycobacterium tuberculosis CYP144: Common themes and lessons learned in the M. tuberculosis P450 enzyme family

Driscoll, Max D.; McLean, Kirsty J.; Cheesman, Myles R.; Jowitt, Thomas A.; Howard, Marjorie; Carroll, Paul; Parish, Tanya; Munro, Andrew W.

doi:10.1016/j.bbapap.2010.05.015

Cited by 24 publications

(43 citation statements)

References 67 publications

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“…When expressed recombinantly in vitro , Cyp142 in addition to Cyp125 catalyze the oxidation of C26 in the side chain of cholesterol. Expression of either cyp125 or cyp142 can support the growth of Δcyp125 Mtb on cholesterol (Driscoll et al, 2011, Johnston et al, 2010). Therefore Cyp142 provides compensatory activity in Mtb H37Rv.…”

Section: Cholesterol Side Chain Metabolismmentioning

confidence: 99%

Pathogen roid rage: Cholesterol utilization byMycobacterium tuberculosis

Wipperman

Sampson

Thomas

2014

Critical Reviews in Biochemistry and Molecular Biology

142

159

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The ability of science and medicine to control the pathogen Mycobacterium tuberculosis (Mtb) requires an understanding of the complex host environment within which it resides. Pathological and biological evidence overwhelmingly demonstrate how the mammalian steroid cholesterol is present throughout the course of infection. Better understanding Mtb requires a more complete understanding of how it utilizes molecules like cholesterol in this environment to sustain the infection of the host. Cholesterol uptake, catabolism, and broader utilization are important for maintenance of the pathogen in the host and it has been experimentally validated to contribute to virulence and pathogenesis. Cholesterol is catabolized by at least three distinct sub-pathways, two for the ring system and one for the side chain, yielding dozens of steroid intermediates with varying biochemical properties. Our ability to control this worldwide infectious agent requires a greater knowledge of how Mtb uses cholesterol to its advantage throughout the course of infection. Herein, the current state of knowledge of cholesterol metabolism by Mtb is reviewed from a biochemical perspective with a focus on the metabolic genes and pathways responsible for cholesterol steroid catabolism.

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Section: Cholesterol Side Chain Metabolismmentioning

confidence: 99%

Pathogen roid rage: Cholesterol utilization byMycobacterium tuberculosis

Wipperman

Sampson

Thomas

2014

Critical Reviews in Biochemistry and Molecular Biology

142

159

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“…Azole drugs, most notably econazole, exhibit antimycobacterial activities against both latent Mtb and multidrug-resistant strains [60, 61], a finding that implicates one or more cytochrome P450 enzymes as targets (Table 1). In this regard, all the Mtb cytochrome P450 enzymes characterized to date, including CYP125 and CYP142, bind azole drugs with low- to submicromolar affinities (Table 1) [62–67]. The crystal structure of CYP125 in complex with econazole, solved at a resolution of 2.2 Å, showed that although the ligand occupies most of the hydrophobic binding pocket, the azole group does not approach the heme iron due to the funnel shape of the active site [55].…”

Section: Cholesterol Catabolism Inhibitorsmentioning

confidence: 99%

Cholesterol catabolism as a therapeutic target in Mycobacterium tuberculosis

Ouellet

Johnston

Montellano

2011

Trends in Microbiology

174

176

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Mycobacterium tuberculosis (Mtb) is an intracellular pathogen that infects 10 million worldwide and kills 2 million people every year. The uptake and utilization of nutrients by Mtb within the host cell is still poorly understood, although lipids play an important role in Mtb persistence. The recent identification of a large regulon of cholesterol catabolic genes suggests that Mtb can use host sterol for infection and persistence. In this review, we report on recent progress in elucidation of the Mtb cholesterol catabolic reactions and their potential utility as targets for tuberculosis therapeutic agents.

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“…It was found that the Mtb CYP144A1 deletion strain was viable and grew in vitro , but that the growth rate of the deletion strain was substantially lower than that of the wild-type Mtb H37Rv. In addition, the CYP144A1 deletion strain was markedly more susceptible to growth inhibition by azole drugs than the wild-type Mtb 22. These data point to important function(s) for CYP144A1 in Mtb , and possibly to its participation in the Mtb stress response mechanisms.…”

Section: Resultsmentioning

confidence: 82%

“…Thus, the prediction of the physiological function of CYP144A1 has been difficult from its genetic context alone22. The nearby gene Rv1771 encodes a L -gulono-1,4-lactone dehydrogenase shown to catalyse the final step required in L -ascorbic acid biosynthesis24, while the Rv1781c gene is predicted to encode a glucanotransferase enzyme (MalQ or amylomaltase)2526.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to the potential of azole drugs as novel therapeutics against Mtb , they are also useful tools for probing the structure and function of the Mtb P450s8. Recently, Mtb CYP144A1, encoded by the H37Rv gene Rv1777 2223, was identified to bind tightly to several azole antifungals, raising the possibility that this P450 could be a target for novel anti-TB drug development.…”

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confidence: 99%

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Structural characterization of CYP144A1 – a cytochrome P450 enzyme expressed from alternative transcripts in Mycobacterium tuberculosis

Chenge

Kavanagh

Driscoll

et al. 2016

Sci Rep

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Mycobacterium tuberculosis (Mtb) causes the disease tuberculosis (TB). The virulent Mtb H37Rv strain encodes 20 cytochrome P450 (CYP) enzymes, many of which are implicated in Mtb survival and pathogenicity in the human host. Bioinformatics analysis revealed that CYP144A1 is retained exclusively within the Mycobacterium genus, particularly in species causing human and animal disease. Transcriptomic annotation revealed two possible CYP144A1 start codons, leading to expression of (i) a “full-length” 434 amino acid version (CYP144A1-FLV) and (ii) a “truncated” 404 amino acid version (CYP144A1-TRV). Computational analysis predicted that the extended N-terminal region of CYP144A1-FLV is largely unstructured. CYP144A1 FLV and TRV forms were purified in heme-bound states. Mass spectrometry confirmed production of intact, His6-tagged forms of CYP144A1-FLV and -TRV, with EPR demonstrating cysteine thiolate coordination of heme iron in both cases. Hydrodynamic analysis indicated that both CYP144A1 forms are monomeric. CYP144A1-TRV was crystallized and the first structure of a CYP144 family P450 protein determined. CYP144A1-TRV has an open structure primed for substrate binding, with a large active site cavity. Our data provide the first evidence that Mtb produces two different forms of CYP144A1 from alternative transcripts, with CYP144A1-TRV generated from a leaderless transcript lacking a 5′-untranslated region and Shine-Dalgarno ribosome binding site.

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Expression and characterization of Mycobacterium tuberculosis CYP144: Common themes and lessons learned in the M. tuberculosis P450 enzyme family

Cited by 24 publications

References 67 publications

Pathogen roid rage: Cholesterol utilization byMycobacterium tuberculosis

Pathogen roid rage: Cholesterol utilization byMycobacterium tuberculosis

Cholesterol catabolism as a therapeutic target in Mycobacterium tuberculosis

Structural characterization of CYP144A1 – a cytochrome P450 enzyme expressed from alternative transcripts in Mycobacterium tuberculosis

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