Study of the preferred modification sites of the quinone methide intermediate resulting from the latent trapping device of the activity probes for hydrolases
Resource
Biochemical & Biophysical Research Communications (326),30–35
Journal
Biochemical & Biophysical Research Communications
Journal Volume
326
Journal Issue
1
Pages
30-35
Date Issued
2005
Date
2005
Author(s)
Lo, Lee-Chiang
Chiang, Ying-Ling
Kuo, Chi-Hsien
Liao, Hsin-Kai
Chen, Yu-Ju
Lin, Jing-Jer
DOI
246246/2006111501265139
Abstract
Use of activity probes has been demonstrated to be a powerful tool in modern chemical proteomic study. Previously we have designed & synthesized a series of mechanism-based activity probes that utilized quinone methide chemistry. Here, we characterized the trend of chemical reactivity for the reactive quinone methide intermediate 3 (QM-3) resulting from the latent trapping device. In a competition assay, the labeling of PTP1B by probe 1a was blocked by externally added cysteine without affecting the catalytic activity of the enzyme. Further sequencing analysis on the trypsin-digested peptides of probe 1a-labeled PTP1B using tandem mass spectrometry revealed that all six cysteine residues of PTP1B are capable of being modified by probe 1a. These results indicated that the sulfhydryl group of cysteine residue is the preferred nucleophile for the reactive QM-3. Our finding provides the first example in understanding the preferred amino acid residues modified by the reactive QM-3, which is also the key structural unit responsible for forming covalent bonds in many biochemical applications.
Subjects
uinone methide
Hydrolases
Protein tyrosine phosphatase
Glucosidase
Cysteine
Labeling
Proteome
Activity probe
Mechanismbased
SDGs
Publisher
Taipei:National Taiwan University Dept Chem Engn
Type
journal article
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