The crystal structure of monoacylglycerol lipase from M. tuberculosis reveals the basis for specific inhibition

Aschauer, P.; Zimmermann, R.; Breinbauer, Rolf; Pavkov‐Keller, Tea; Oberer, Monika

doi:10.1038/s41598-018-27051-7

Cited by 25 publications

(35 citation statements)

References 43 publications

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“…The homology model of PFL was generated using Swiss-Model 2 ( Biasini et al, 2014 ) with the crystal structure of Pseudomonas aeruginosa lipase (PDB ID: 1EX9) as the template ( Nardini et al, 2000 ; Aschauer et al, 2018 ). VERIFY-3D was used to determine the compatibility of the atomic model (3D) with the amino acid sequence (1D) 3 ( Lüthy et al, 1992 ).…”

Section: Methodsmentioning

confidence: 99%

Directed Evolution of Pseudomonas fluorescens Lipase Variants With Improved Thermostability Using Error-Prone PCR

Guan¹,

Gao²,

Li³

et al. 2020

Front. Bioeng. Biotechnol.

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Lipases catalyze the hydrolysis of fats and oils, and have been widely used in various industrial fields. However, bacterial lipases have a lower thermostability in industrial processes, which was a limiting factor in their industrial application. In this study, we obtained an improve variant of Pseudomonas fluorescens lipase (PFL) with enhanced thermostability using classical error-prone PCR. Wild-type PFL showed an optimal temperature and pH of 50°C and pH 7.5, respectively. Due to the low thermostability of PFL, a library containing over 3000 individual mutants as constructed using error-prone PCR. Screening for thermotolerance yielded the mutants L218P and P184C/M243C with T m values of 62.5 and 66.0°C, which was 2.5 and 6°C higher than that of the WT, respectively. The combination of the two mutants (P184C/M243C/L218P) resulted in an approximately additive effect with a T m value of 68.0°C. Although the increase of T m was not substantial, the mutant also had dramatically increased methanol tolerance. Structural analysis revealed that the introduction of a disulfide bond between P184C and M243C and the substitution of Pro to reduce the flexibility of a loop increased the thermostability of PFL, which provides a theoretical foundation for improving the thermostability and methanol tolerance of lipase family I.1 to resist the harsh conditions of industrial processes.

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Section: Methodsmentioning

confidence: 99%

Directed Evolution of Pseudomonas fluorescens Lipase Variants With Improved Thermostability Using Error-Prone PCR

Guan¹,

Gao²,

Li³

et al. 2020

Front. Bioeng. Biotechnol.

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“…A DALI search retrieved hMGL as the first hit [ 7 , 25 , 26 , 34 ]. Other close structural homologs of PFL include mtMGL and Saccharomyces cerevisiae (scMGL) monoglyceride lipases [ 8 , 35 ]. Despite the relatively low sequence identity with these lipases (32%, 37%, and 23% identity with hMGL, mtMGL, and scMGL, respectively) the enzymes retain a striking conservation of structural elements, which are presumed to be of critical importance to support the activity ( Figure S1 ).…”

Section: Resultsmentioning

confidence: 99%

“…For instance, since the enzyme was shown to constitute a key actor of the endocannabinoid signaling system, a great deal of attention was devoted to the human monoglyceride lipase (hMGL), which is now considered as a promising drug target for the treatment of several disorders including cancer and inflammatory diseases [ 6 , 7 ]. Homologs of hMGL were also identified in bacteria, yeast, and recently, mycobacterium tuberculosis MGL (mtMGL) inhibition emerged as a new therapeutic strategy to treat tuberculosis [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

Structure and Dynamics of an Archeal Monoglyceride Lipase from Palaeococcus ferrophilus as Revealed by Crystallography and In Silico Analysis

Labar¹,

Brandt²,

Flaba³

et al. 2021

Biomolecules

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The crystallographic analysis of a lipase from Palaeococcus ferrophilus (PFL) previously annotated as a lysophospholipase revealed high structural conservation with other monoglyceride lipases, in particular in the lid domain and substrate binding pockets. In agreement with this observation, PFL was shown to be active on various monoacylglycerols. Molecular Dynamics (MD) studies performed in the absence and in the presence of ligands further allowed characterization of the dynamics of this system and led to a systematic closure of the lid compared to the crystal structure. However, the presence of ligands in the acyl-binding pocket stabilizes intermediate conformations compared to the crystal and totally closed structures. Several lid-stabilizing or closure elements were highlighted, i.e., hydrogen bonds between Ser117 and Ile204 or Asn142 and its facing amino acid lid residues, as well as Phe123. Thus, based on this complementary crystallographic and MD approach, we suggest that the crystal structure reported herein represents an open conformation, at least partially, of the PFL, which is likely stabilized by the ligand, and it brings to light several key structural features prone to participate in the closure of the lid.

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“…Furthermore, disruption of the ortholog of Rv0183 in Mycobacterium smegmatis has been shown to impact drug susceptibility ( Dhouib et al, 2010 ). These features make these enzymes, along with their accessible location, suitable targets for anti-tubercular drugs, which are currently being pursued ( Aschauer et al, 2018 , Point et al, 2013 , Point et al, 2012 , Saravanan et al, 2012 ). As we have proposed that Rv0183 is involved in the activation of the HTB compounds, inhibition of this enzyme would be undesirable in the context of the potential treatment with compounds of this class.…”

Section: Discussionmentioning

confidence: 99%

Anti-tubercular derivatives of rhein require activation by the monoglyceride lipase Rv0183

Abrahams

et al. 2020

The Cell Surface

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The emergence and perseverance of drug resistant strains of Mycobacterium tuberculosis ( Mtb ) ensures that drug discovery efforts remain at the forefront of tuberculosis research. There are numerous different approaches that can be employed to lead to the discovery of anti-tubercular agents. In this work, we endeavored to optimize the anthraquinone chemical scaffold of a known drug, rhein, converting it from a compound with negligible activity against Mtb , to a series of compounds with potent activity. Two compounds exhibited low toxicity and good liver microsome stability and were further progressed in attempts to identify the biological target. Whole genome sequencing of resistant isolates revealed inactivating mutations in a monoglyceride lipase. Over-expression trials and an enzyme assay confirmed that the designed compounds are prodrugs, activated by the monoglyceride lipase. We propose that rhein is the active moiety of the novel compounds, which requires chemical modifications to enable access to the cell through the extensive cell wall structure. This work demonstrates that re-engineering of existing antimicrobial agents is a valid method in the development of new anti-tubercular compounds.

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The crystal structure of monoacylglycerol lipase from M. tuberculosis reveals the basis for specific inhibition

Cited by 25 publications

References 43 publications

Directed Evolution of Pseudomonas fluorescens Lipase Variants With Improved Thermostability Using Error-Prone PCR

Directed Evolution of Pseudomonas fluorescens Lipase Variants With Improved Thermostability Using Error-Prone PCR

Structure and Dynamics of an Archeal Monoglyceride Lipase from Palaeococcus ferrophilus as Revealed by Crystallography and In Silico Analysis

Anti-tubercular derivatives of rhein require activation by the monoglyceride lipase Rv0183

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