Molecular Basis of Chiral Acid Recognition by <i>Candida rugosa</i> Lipase: X‐Ray Structure of Transition State Analog and Modeling of the Hydrolysis of Methyl 2‐Methoxy‐2‐phenylacetate

Colton, Ian J.; Yin, Tyler; Grochulski, P.; Kazlauskas, Romas J.

doi:10.1002/adsc.201100459

Cited by 24 publications

(22 citation statements)

References 45 publications

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“…We synthesized p- nitrophenol esters of alkyl phosphonic acids and used them as substrate analogs with different alkyl chain lengths. These compounds react irreversibly with the nucleophilic serine of α/β-hydrolases and are used frequently to capture the tetrahedral reaction intermediates of lipases ( 41 – 45 ). Consequently, the inhibitor complexes go one step further along the reaction trajectory compared with the D196N 1-LG complex; Ser-97 exerted its nucleophilic attack on the phosphorus-atom of the MG-mimicking ligand, and then the reaction is stalled.…”

Section: Resultsmentioning

confidence: 99%

Conformational Plasticity and Ligand Binding of Bacterial Monoacylglycerol Lipase

Rengachari

Aschauer

Schittmayer

et al. 2013

Journal of Biological Chemistry

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Background: Monoacylglycerol lipases hydrolyze monoacylglycerols into free fatty acids and glycerols.Results: Crystal structures provide the structural basis for conformational plasticity and ligand binding of a monoacylglycerol lipase.Conclusion: Hallmark features responsible for substrate binding and selectivity are conserved across species.Significance: The first structures of a monoacylglycerol lipase in complex with substrate analogs are presented.

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Section: Resultsmentioning

confidence: 99%

Conformational Plasticity and Ligand Binding of Bacterial Monoacylglycerol Lipase

Rengachari

Aschauer

Schittmayer

et al. 2013

Journal of Biological Chemistry

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“…In the Protein Data Bank (PDB), there are 12 structures available for lipase B from Candida antarctica (CAL-B): 1TCA, 1TCB and 1TCC (Uppenberg et al, 1994), 1LBS and 1LBT (Uppenberg et al, 1995), 3ICV and 3ICW (Qian et al, 2009), 4K5Q, 4K6G, 4K6H, and 4K6K (Xie et al, 2014), and 3W9B (Kim et al, 2014). For Candida rugosa lipase (CRL), there are 9 structures: 1CRL (Grochulski et al, 1993), 1GZ7 (Mancheno et al, 2003), 1LPM and 1LPS (Cygler et al, 1994), 1LPN, 1LPO, and 1LPP (Grochulski et al, 1994a), 1TRH (Grochulski et al, 1994b), and 3RAR (Colton et al, 2011). For lipase PS Amano, there is one PDB entry, 1OIL (Kim et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Crystal and molecular structure of hexagonal form of lipase B from Candida antarctica.

Strzelczyk¹,

Bujacz²,

Kiełbasiński³

et al. 2015

Acta Biochim Pol

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During crystallization screenings of commercially available hydrolytic enzymes, the new, hexagonal crystal form of CAL-B, has been discovered and hereby reported. The NAG molecules, which were closing the glycosylation site in the orthorhombic form, in hexagonal structure make the glycosylation site open. It is unknown whether the opening and closing of the glycosylation site by the 'lid' NAG molecules, could be related to the opening and closing of the active center of the enzyme upon substrate binding and product release.

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“…The fourth orientation is the umbrella-like inversion orientation. [1,41,42] This orientation is a mirrorimage reflection, but the mirror plane lies not between the enantiomers as is customary, but cuts through the molecule near the stereocenter perpendicular to the C À H bond. This reflection moves the hydrogen substituent to a new, previously unoccupied, region, while leaving the remaining three substituents in approximately the same location.…”

Section: Orientation Of Slow-reacting Enantiomersmentioning

confidence: 96%

“…The primary alcohol substrates are flexible and can adjust to the binding site, but the placement of the stereocenter within a ring limits possible adjustments. Both X-ray crystal structures of enantiomers bound to enzymes [1] and molecular modeling [41,42] show that the umbrella-like orientation is common and must be considered for the slow enantiomer. Previous modeling rarely considered this possibility.…”

Section: Orientation Of Slow-reacting Enantiomersmentioning

confidence: 99%

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Molecular Basis for the Enantio‐ and Diastereoselectivity of Burkholderia cepacia Lipase toward γ‐Butyrolactone Primary Alcohols

Eum

Kazlauskas

2014

Adv Synth Catal

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Burkholderia cepacia lipase (BCL) shows high enantioselectivity toward chiral primary alcohols, but this enantioselectivity is often unpredictable, especially for substrates that contain an oxygen at the stereocenter. For example, BCL resolves bsubstituted-g-acetyloxymethyl-g-butyrolactones (acetates of a chiral primary alcohol) by hydrolysis of the acetate, but the enantioselectivity varies with the nature and orientation of the b-alkyl substituent. BCL favors the (R)-primary alcohol when the balkyl substituent is hydrogen (E = 30) or trans methyl (E = 38), but the (S)-primary alcohol when it is cis methyl (E = 145). To rationalize this unusual selectivity, we used a combination of experiments to show the importance of polar interactions and modeling to reveal differences in orientations of the enantiomers. Removal of either the lactone carbonyl in the substrate or the polar side chains in the enzyme by using a related enzyme without these side chains decreased the enantioselectivity at least four-fold. Modeling revealed that the slow enantiomers do not bind by exchanging the location of two substituents relative to the fast enantiomer. Instead, three substituents remain in the same region, but the fourth substituent, hydrogen, inverts to a new location, like an umbrella in a strong wind. In this orientation the favored stereoisomers have similar shapes, thus accounting for the unusual stereoselectivity. The ratio of catalytically productive orientations for the fast vs. slow enantiomers in a molecular dynamic simulation correlated (R 2 = 0.82) with the degree of enantioselectivity including the case where the enantioselectivity reversed. Weighting this ratio by the ratio of Hbonds in the polar interaction to account for different binding strengths improved the correlation with the measured enantioselectivity to R 2 = 0.97. The modeling identifies key interactions responsible for high enantioselectivity in this class of substrates.

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Molecular Basis of Chiral Acid Recognition by Candida rugosa Lipase: X‐Ray Structure of Transition State Analog and Modeling of the Hydrolysis of Methyl 2‐Methoxy‐2‐phenylacetate

Cited by 24 publications

References 45 publications

Conformational Plasticity and Ligand Binding of Bacterial Monoacylglycerol Lipase

Conformational Plasticity and Ligand Binding of Bacterial Monoacylglycerol Lipase

Crystal and molecular structure of hexagonal form of lipase B from Candida antarctica.

Molecular Basis for the Enantio‐ and Diastereoselectivity of Burkholderia cepacia Lipase toward γ‐Butyrolactone Primary Alcohols

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