Document Detail


Modulation of infectivity in phage display as a tool to determine the substrate specificity of proteases.
MedLine Citation:
PMID:  16642518     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Proteases play an important role in human and animal diseases. Rapid determination of substrate specificity is possible through the use of substrate phage display; however, current methods possess several drawbacks. They require phage-immobilization and cannot be used for infectivity-destroying or affinity tag-destroying proteases; this can make entire libraries useless. To overcome these limitations, here we introduce infectivity-modulated phage display (IMOP). IMOP uses a protease-resistant and infectivity-reducing tag fused to substrate-displaying polyvalent phages, and the specific cleavage of the substrate increases the infectivity of the phages by releasing the infectivity-reducing tag. The resulting phages were first tested with the infectivity-destroying detergent protease subtilisin; this resulted in a highly specific substrate at a 200-fold enrichment. In a second example, the protease ompT was used and led to an enrichment of the known double-arginine motif. The IMOP system thus substantially improves and simplifies previous systems.
Authors:
Javier F Chaparro-Riggers; Roland Breves; Karl-Heinz Maurer; Uwe Bornscheuer
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Chembiochem : a European journal of chemical biology     Volume:  7     ISSN:  1439-4227     ISO Abbreviation:  Chembiochem     Publication Date:  2006 Jun 
Date Detail:
Created Date:  2006-05-31     Completed Date:  2006-11-08     Revised Date:  2012-07-11    
Medline Journal Info:
Nlm Unique ID:  100937360     Medline TA:  Chembiochem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  965-70     Citation Subset:  IM    
Affiliation:
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, Greifswald University, Soldmannstrasse 16, 17487 Greifswald, Germany.
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MeSH Terms
Descriptor/Qualifier:
Peptide Hydrolases / chemistry*
Peptide Library*
Serine Endopeptidases / chemistry,  metabolism
Substrate Specificity
Subtilisin / chemistry,  metabolism
Chemical
Reg. No./Substance:
0/Peptide Library; EC 3.4.-/Peptide Hydrolases; EC 3.4.21.-/Serine Endopeptidases; EC 3.4.21.62/Subtilisin; EC 3.4.23.49/omptin outer membrane protease

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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