Conformational Plasticity and Ligand Binding of Bacterial Monoacylglycerol Lipase

Rengachari, Srinivasan; Aschauer, P.; Schittmayer, Matthias; Mayer, Nicole; Gruber, Karl; Breinbauer, Rolf; Birner‐Gruenberger, Ruth; Dreveny, Ingrid; Oberer, Monika

doi:10.1074/jbc.m113.491415

Cited by 45 publications

(54 citation statements)

References 42 publications

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“…The orientation of the polar head of monoacylglycerols is well established based on earlier modeling studies as well as recent experimental structures of bacterial MAGL in various conformations (Bertrand et al, 2010;Rengachari et al, 2013). To gain further understanding on the binding properties in the ground state, a series of unconstrained molecular dynamic simulations were calculated for WT and mutant complexes with selected docking poses of 2-and 1-AG.…”

Section: Structural Determinants Of Magl For Substrate Preferencementioning

confidence: 99%

Mutation of Cys242 of Human Monoacylglycerol Lipase Disrupts Balanced Hydrolysis of 1- and 2-Monoacylglycerols and Selectively Impairs Inhibitor Potency

et al. 2013

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Considerable progress has been made in recent years in developing selective, potent monoacylglycerol lipase (MAGL) inhibitors. In the investigations of measures to inhibit this enzyme, less attention has been paid to improving our understanding of its catalytic mechanisms or substrate preferences. In our study, we used site-directed mutagenesis, and we show via versatile activity assays combined with molecular modeling that Cys242 and Tyr194, the two opposing amino acid residues in the catalytic cavity of MAGL, play important roles in determining the rate and the isomer preferences of monoacylglycerol hydrolysis. In contrast to wild-type enzymes that hydrolyzed 1-and 2-monoacylglycerols at similar rates, mutation of Cys242 to alanine caused a significant reduction in overall activity (maximal velocity, V max ), particularly skewing the balanced hydrolysis of isomers to favor the 2-isomer. Molecular modeling studies indicate that this was caused by structural features unfavorable toward 1-isomers as well as impaired recognition of OH-groups in the glycerol moiety. Direct functional involvement of Cys242 in the catalysis was found unlikely due to the remote distance from the catalytic serine. Unlike C242A, mutation of Tyr194 did not bias the hydrolysis of 1-and 2-monoacylglycerols but significantly compromised overall activity. Finally, mutation of Cys242 was also found to impair inhibition of MAGL, especially that by fluorophosphonate derivatives (13-to 63-fold reduction in potency). Taken together, this study provides new experimental and modeling insights into the molecular mechanisms of MAGLcatalyzed hydrolysis of the primary endocannabinoid 2-arachidonoylglycerol and related monoacylglycerols.

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Section: Structural Determinants Of Magl For Substrate Preferencementioning

confidence: 99%

Mutation of Cys242 of Human Monoacylglycerol Lipase Disrupts Balanced Hydrolysis of 1- and 2-Monoacylglycerols and Selectively Impairs Inhibitor Potency

et al. 2013

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“…Additionally, a number of crystal structures have been produced for the enzyme-ligand complexes. 154, 155 Our involvement with MGL stemmed from an interest in obtaining detailed information on its structure and function beyond what was available in the literature, and in using it to design and synthesize novel inhibitors covering a broad spectrum of potency, reversibility, and selectivity profiles. Additionally, we were interested in exploring MGL species differences with the intent of developing novel successful inhibitors that were equally potent in human and rodent enzymes to ensure that if successful, these compounds could be advanced beyond the preclinical stage.…”

Section: Endocannabinoid Deactivating Enzymesmentioning

confidence: 99%

2012 Division of Medicinal Chemistry Award Address. Trekking the Cannabinoid Road: A Personal Perspective

Makriyannis

2014

J. Med. Chem.

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My involvement with the field of cannabinoids spans close to three decades, and covers a major part of my scientific career. It also reflects the robust progress in this initially largely unexplored area of biology. During this period of time, I have witnessed the growth of modern cannabinoid biology, starting from the discovery of its two receptors and followed by the characterization of its endogenous ligands and the identification of the enzyme systems involved in their biosynthesis and biotransformation. I was fortunate enough to start at the beginning of this new era and participate in a number of the new discoveries. It has been a very exciting journey. By covering some key aspects of my work during this period of “modern cannabinoid research,” this perspective, in part historical, intends to give an account of how the field grew, the key discoveries, and the most promising directions for the future.

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“…H257 have been determined to date (Bertrand et al, 2010;Labar et al, 2010;Schalk-Hihi et al, 2011;Rengachari et al, 2012Rengachari et al, , 2013. The structures revealed an / hydrolase fold with a dynamic yet topologically conserved cap region.…”

Section: Introductionmentioning

confidence: 99%

Purification, crystallization and preliminary X-ray diffraction analysis of a soluble variant of the monoglyceride lipase Yju3p from the yeastSaccharomyces cerevisiae

et al. 2015

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The protein Yju3p is the orthologue of monoglyceride lipases in the yeastSaccharomyces cerevisiae. A soluble variant of this lipase termed s-Yju3p (38.3 kDa) was generated and purified to homogeneity by affinity and size-exclusion chromatography. s-Yju3p was crystallized in a vapour-diffusion setup at 293 K and a complete data set was collected to 2.4 Å resolution. The crystal form was orthorhombic (space groupP212121), with unit-cell parametersa= 77.2,b= 108.6,c= 167.7 Å. The asymmetric unit contained four molecules with a solvent content of 46.4%.

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Conformational Plasticity and Ligand Binding of Bacterial Monoacylglycerol Lipase

Cited by 45 publications

References 42 publications

Mutation of Cys242 of Human Monoacylglycerol Lipase Disrupts Balanced Hydrolysis of 1- and 2-Monoacylglycerols and Selectively Impairs Inhibitor Potency

Mutation of Cys242 of Human Monoacylglycerol Lipase Disrupts Balanced Hydrolysis of 1- and 2-Monoacylglycerols and Selectively Impairs Inhibitor Potency

2012 Division of Medicinal Chemistry Award Address. Trekking the Cannabinoid Road: A Personal Perspective

Purification, crystallization and preliminary X-ray diffraction analysis of a soluble variant of the monoglyceride lipase Yju3p from the yeastSaccharomyces cerevisiae

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