Emergence of a novel highly specific and catalytically efficient enzyme from a naturally promiscuous glutathione transferase

Blikstad, Cecilia; Shokeer, Abeer; Kurtovic, Sanela; Mannervik, Bengt

doi:10.1016/j.bbagen.2008.07.007

Cited by 17 publications

(24 citation statements)

References 29 publications

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“…Additional studies with the aforementioned GIMFhelix mutant further established the relationship between the structural determinants that contribute to the distinct specificities of alpha-class GSTs (Balogh et al, 2009; Blikstad et al, 2008; Nilsson et al, 2000). Because a number of the targeted residues comprise the α9-helix, the entire C-terminus (residues 208–222) was exchanged with that of GSTA4-4 in the final GSTA1-1 GIMFhelix mutant designed by Mannervik and co-workers Nilsson et al, 2000.…”

Section: Structural Characterizationsmentioning

confidence: 95%

“…Rationally engineered mutants of GSTA1-1, including the elegantly redesigned GSTA1-1 “GIMFhelix” mutant, which contains amino-acid substitutions representing 6% of the sequence (A12G/L107I/L108M/V111F/A1 208–222 A4), have also uncovered structural elements important within the context of high activity toward alkenals (Blikstad et al, 2008; Nilsson et al, 2000; Babbitt, 2000). Although it is still less efficient than GSTA4-4, the GIMFhelix mutant is remarkably more active with HNE than GSTA1-1.…”

Section: Detoxification Of Hnementioning

confidence: 99%

“…The GSTA4-4 isoform is known to be selective for the GSH conjugation of LPO products and is well-recognized for its catalytic efficiency with HNE (Board, 1998; Hubatsch et al, 1998). Structurally, this isoform consists of a rigid scaffold preorganized for HNE metabolism, whereas the structural homolog GSTA1-1, which has low activity toward HNE, utilizes a conformationally flexible scaffold to accomplish promiscuous catalysis with diverse substrates, such as 1-chloro-2,4-dinitrobenzene (CDNB) and Δ 5 -androstene-3,17-dione (Balogh et al, 2010; Bruns et al, 1999; Hou et al, 2007; Blikstad et al, 2008). However, in addition to directly metabolizing HNE, some of the alpha-class GSTs can also provide antioxidant protection by attenuating HNE formation.…”

Section: Introductionmentioning

confidence: 99%

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Interactions of glutathione transferases with 4-hydroxynonenal

Balogh

Atkins

2011

Drug Metabolism Reviews

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Electrophilic products of lipid peroxidation are important contributors to the progression of several pathological states. The prototypical α,β–unsaturated aldehyde, 4-hydroxynonenal (HNE), triggers cellular events associated with oxidative stress, which can be curtailed by the glutathione-dependent elimination of HNE. The glutathione transferases (GSTs) are a major determinate of the intracellular concentration of HNE and can influence susceptibility to toxic effects, particularly when HNE and GST levels are altered in disease states. In this article, we provide a brief summary of the cellular effects of HNE, followed by a review of its GST-catalyzed detoxification, with an emphasis on the structural attributes that play an important role in the interactions with alpha-class GSTs. Some of the key determining characteristics that impart high alkenal activity reside in the unique C-terminal interactions of the GSTA4-4 enzyme. Studies encompassing both kinetic and structural analyses of related isoforms will be highlighted, with additional attention to stereochemical aspects that demonstrate the capacity of GSTA4-4 to detoxify both enantiomers of the biologically relevant racemic mixture while generating a select set of diastereomeric products with subsequent implications. A summary of the literature that examines the interplay between GSTs and HNE in model systems relevant to oxidative stress will also be discussed to demonstrate the magnitude of importance of GSTs in the overall detoxification scheme.

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Section: Structural Characterizationsmentioning

confidence: 95%

Section: Detoxification Of Hnementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interactions of glutathione transferases with 4-hydroxynonenal

Balogh

Atkins

2011

Drug Metabolism Reviews

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“…GSTA4-4 has a static C-terminal helix localized along one edge of the active site that does not undergo any rearrangement upon binding substrate (31,32). The GSTA1-1/GSTA4-4 pair has been a powerful model for engineering substrate specificity and understanding its structural basis in GSTA4-4 (5,12,33,34), but the molecular basis for the contrasting promiscuity of GSTA1-1 is not understood in any thermodynamic context. Therefore, we have performed scanning calorimetry with these GST isoforms and analyzed the results with a modified Landau theory of phase transitions, as described for protein folding (35,36).…”

mentioning

confidence: 99%

Ensemble Perspective for Catalytic Promiscuity

Honaker

Acchione

Sumida

et al. 2011

Journal of Biological Chemistry

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Background: Catalytic promiscuity is common, but its molecular basis is poorly understood. Results: Differential scanning calorimetry reveals the local active site conformational landscape of a promiscuous detoxification enzyme, glutathione transferase A1-1, as "smooth" and heterogeneous. Conclusion: Facile conformational exchange facilitates substrate promiscuity. Significance: The results provide the first thermodynamic basis for catalytic promiscuity.

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“…The ␣-class GST family includes the isoforms GSTA1-1 and A4 -4, which are highly promiscuous and highly substrate specific, respectively, despite their high sequence similarity and nearly superimposable structures (Fig. 2, a and b) (12,18,19). Whereas GSTA1-1 is an archetypal promiscuous detoxication enzyme, GSTA4 -4 has great specificity for straight chain unsaturated lipid aldehydes, about nine carbons in length, that result from lipid peroxidation (20).…”

mentioning

confidence: 99%

Enzymatic Detoxication, Conformational Selection, and the Role of Molten Globule Active Sites

Honaker

Acchione

Zhang

et al. 2013

Journal of Biological Chemistry

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Background: It is unknown whether enzyme promiscuity is achieved by induced fit or conformational selection. Results: Pre-steady state kinetic and thermodynamic analysis of promiscuous enzymes indicates substrate-dependent conformational selection of substrate-free ensembles. Conclusion: Catalytic promiscuity correlates with thermodynamic parameters of substrate-free enzyme conformation. Significance: These results are the first to demonstrate that molten globule active sites facilitate conformational selection.

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Emergence of a novel highly specific and catalytically efficient enzyme from a naturally promiscuous glutathione transferase

Cited by 17 publications

References 29 publications

Interactions of glutathione transferases with 4-hydroxynonenal

Interactions of glutathione transferases with 4-hydroxynonenal

Ensemble Perspective for Catalytic Promiscuity

Enzymatic Detoxication, Conformational Selection, and the Role of Molten Globule Active Sites

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