Trypsin display on the surface of bacteriophage.
The gene III and VIII-encoded coat proteins (pIII and pVIII) from bacteriophage M13 have been fused to the C terminus of the serine protease, trypsin (Tsn). The genes encoding the fusions were then inserted directly into M13mp18 to create vectors which expressed both the Tsn-coat protein hybrids and the wild-type (wt) coat proteins. Immunoblot analysis confirmed that the bacteriophage express Tsn on their surface. Isolated fusion phage possess kinetic parameters which approximate those of the wt enzyme. An endogenous Escherichia coli protease inhibitor, ecotin, copurifies with the Tsn phage. Immobilized ecotin can be used to selectively bind bacteriophage which express Tsn::pIII fusion proteins.
Corey
D R
DR
Department of Pharmaceutical Chemistry, University of California San Francisco 94143.
Shiau
A K
AK
Yang
Q
Q
Janowski
B A
BA
Craik
C S
CS
D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification; D017104:Bacteriophage M13 / Q000235:genetics*; D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D036022:Capsid Proteins; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D005814:Genes, Viral; D005822:Genetic Vectors; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D033903:Periplasmic Proteins; D010957:Plasmids; D016133:Polymerase Chain Reaction; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D015183:Restriction Mapping; D014357:Trypsin / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D014361:Trypsin Inhibitors / Q000302:isolation & purification; D014760:Viral Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D036022:Capsid Proteins; D004268:DNA-Binding Proteins; C039076:Eco protein, E coli; D029968:Escherichia coli Proteins; D009838:Oligodeoxyribonucleotides; D033903:Periplasmic Proteins; D011993:Recombinant Fusion Proteins; D014361:Trypsin Inhibitors; D014760:Viral Fusion Proteins; D014357:Trypsin