Use of benzyl mercaptan for direct preparation of long polypeptide benzylthio esters as substrates of subtiligase

Details

Serval ID
serval:BIB_D46F3CA5781D
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Use of benzyl mercaptan for direct preparation of long polypeptide benzylthio esters as substrates of subtiligase
Journal
Biochemical and Biophysical Research Communications
Author(s)
Welker  E., Scheraga  H. A.
ISSN
0006-291X (Print)
Publication state
Published
Issued date
01/1999
Volume
254
Number
1
Pages
147-51
Notes
Journal Article
Research Support, U.S. Gov't, P.H.S. --- Old month value: Jan 8
Abstract
Subtiligase, a double mutant of subtilisin, has been shown to be capable of joining together two unprotected peptide fragments, namely an activated peptide ester and a second peptide with a free N-terminal amino group. Inside cells, inteins are know to join peptide chains (exteins) by self-extrusion. The SC VMA1 intein was modified to undergo only in vitro N-terminal cleavage in the presence of small nucleophilic compounds, releasing the N-terminal extein. With a proper choice of the nucleophilic compounds it is shown that it is possible to generate long polypeptides, by molecular biology expression, with such an attached reactive ester which is an excellent substrate of the enzyme, subtiligase. This approach can successfully extend the current limit of the subtiligase-catalyzed fragment condensation method as well as provide another application of the recently discovered intein chemistry.
Keywords
*Benzyl Compounds Escherichia coli Mutation Peptide Synthases/chemistry/*metabolism Peptides/*chemistry/metabolism Substrate Specificity Subtilisins/chemistry/genetics/*metabolism *Sulfhydryl Compounds
Pubmed
Web of science
Create date
24/01/2008 14:40
Last modification date
20/08/2019 15:54
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