The mechanism of the amidases: Mutating the glutamate adjacent to the catalytic triad inactivates the enzyme due to substrate mispositioning

Brandon W. Weber; Serah W. Kimani; Arvind Varsani; Donald A. Cowan; Roger Hunter; Gerhard A. Venter; James C. Gumbart; B. Trevor Sewell

doi:10.1074/jbc.M113.503284

The mechanism of the amidases: Mutating the glutamate adjacent to the catalytic triad inactivates the enzyme due to substrate mispositioning

Brandon W. Weber, Serah W. Kimani, Arvind Varsani, Donald A. Cowan, Roger Hunter, Gerhard A. Venter, James C. Gumbart, B. Trevor Sewell

Research output: Contribution to journal › Article › peer-review

29 Scopus citations

Abstract

Background:A cysteine, a glutamic acid, and a lysine are the well known amidase catalytic residues. Results:Mutating the neighboring, structurally conserved Glu-142 inactivates the enzyme, but the active site cysteine still reacts with acrylamide via its double bond. Conclusion:Glu-142 positions the amide for productive nucleophilic attack by the cysteine. Significance:An intact catalytic tetrad is required for amidase activity.

Original language	English (US)
Pages (from-to)	28514-28523
Number of pages	10
Journal	Journal of Biological Chemistry
Volume	288
Issue number	40
DOIs	https://doi.org/10.1074/jbc.M113.503284
State	Published - Oct 4 2013
Externally published	Yes

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Cell Biology

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title = "The mechanism of the amidases: Mutating the glutamate adjacent to the catalytic triad inactivates the enzyme due to substrate mispositioning",

abstract = "Background:A cysteine, a glutamic acid, and a lysine are the well known amidase catalytic residues. Results:Mutating the neighboring, structurally conserved Glu-142 inactivates the enzyme, but the active site cysteine still reacts with acrylamide via its double bond. Conclusion:Glu-142 positions the amide for productive nucleophilic attack by the cysteine. Significance:An intact catalytic tetrad is required for amidase activity.",

author = "Weber, {Brandon W.} and Kimani, {Serah W.} and Arvind Varsani and Cowan, {Donald A.} and Roger Hunter and Venter, {Gerhard A.} and Gumbart, {James C.} and Sewell, {B. Trevor}",

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doi = "10.1074/jbc.M113.503284",

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T1 - The mechanism of the amidases

T2 - Mutating the glutamate adjacent to the catalytic triad inactivates the enzyme due to substrate mispositioning

AU - Weber, Brandon W.

AU - Kimani, Serah W.

AU - Varsani, Arvind

AU - Cowan, Donald A.

AU - Hunter, Roger

AU - Venter, Gerhard A.

AU - Gumbart, James C.

AU - Sewell, B. Trevor

PY - 2013/10/4

Y1 - 2013/10/4

N2 - Background:A cysteine, a glutamic acid, and a lysine are the well known amidase catalytic residues. Results:Mutating the neighboring, structurally conserved Glu-142 inactivates the enzyme, but the active site cysteine still reacts with acrylamide via its double bond. Conclusion:Glu-142 positions the amide for productive nucleophilic attack by the cysteine. Significance:An intact catalytic tetrad is required for amidase activity.

AB - Background:A cysteine, a glutamic acid, and a lysine are the well known amidase catalytic residues. Results:Mutating the neighboring, structurally conserved Glu-142 inactivates the enzyme, but the active site cysteine still reacts with acrylamide via its double bond. Conclusion:Glu-142 positions the amide for productive nucleophilic attack by the cysteine. Significance:An intact catalytic tetrad is required for amidase activity.

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The mechanism of the amidases: Mutating the glutamate adjacent to the catalytic triad inactivates the enzyme due to substrate mispositioning

Abstract

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