An Acyloxymethyl Ketone-Based Probe to Monitor the Activity of Glutathionylspermidine Amidase in Escherichia coli

Cellular redox conditions affect Gsp amidase activity in Escherichia coli. Guided by the structure and catalytic mechanism of the amidase, we designed and synthesized an acyloxymethyl ketone‐based activity probe containing a biotin handle. This probe was used to monitor Gsp amidase activity in E. co...

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Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2011-10, Vol.12 (15), p.2306-2309
Main Authors: Chong, Cheong-Meng, Gao, Shijay, Chiang, Bing-Yu, Hsu, Wen-Hung, Lin, Ting-Chien, Chen, Tzu-Chieh, Lin, Chun-Hung
Format: Article
Language:English
Subjects:
activity-based probes
acyloxymethyl ketone
Amidohydrolases - metabolism
Binding Sites
chap domain
Enzyme Assays - methods
Escherichia coli - enzymology
Glutathione - analogs & derivatives
Glutathione - metabolism
glutathionylspermidine amidase
Ketones - chemistry
Ketones - metabolism
Models, Molecular
Oxidation-Reduction
redox chemistry
Spermidine - analogs & derivatives
Spermidine - metabolism
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Summary:Cellular redox conditions affect Gsp amidase activity in Escherichia coli. Guided by the structure and catalytic mechanism of the amidase, we designed and synthesized an acyloxymethyl ketone‐based activity probe containing a biotin handle. This probe was used to monitor Gsp amidase activity in E. coli lysates that had been subjected to oxidative or methylglyoxal‐induced stress.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.201100417