Journal
CHEMBIOCHEM
Volume 7, Issue 5, Pages 834-838Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.200500427
Keywords
combinatorial libraries; peptides; phage display; protease substrates; proteins
Funding
- NCRR NIH HHS [RR19325] Funding Source: Medline
- NIAID NIH HHS [1-U54-AI057153] Funding Source: Medline
- NINDS NIH HHS [NS39892] Funding Source: Medline
- PHS HHS [C76 HF03610-01-00] Funding Source: Medline
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We describe a bacteriophage M13 substrate library encoding the AviTag (BirA substrate) and combinatorial heptamer peptides displayed at the N terminus of the mature form of capsid protein III. Phages are biotinylated efficiently (>= 50 %) when grown in E. coli cells coexpressing BirA, and such viral particles can be immobilized on a streptavidin-coated support and released by protease cleavage within the combinotorial peptide. We have used this library to map the specificity of human Factor Xa and a neuropeptidase, neurolysin (EC3.4.24.16). Validation by analysis of isolated peptide substrates has revealed that neurolysin recognizes the motif hydrophobic-X-Pro-Arg-hydrophobic, where Arg-hydrophobic is the scissile bond.
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