Trypsin display on the surface of bacteriophage

David R. Corey; Andrew K. Shiau; Yang Qing; Bethany A. Janowski; Charles S. Craik

doi:10.1016/0378-1119(93)90163-W

Trypsin display on the surface of bacteriophage

David R. Corey, Andrew K. Shiau, Yang Qing, Bethany A. Janowski, Charles S. Craik

Pharmacology

Research output: Contribution to journal › Article › peer-review

84 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	129-134
Number of pages	6
Journal	Gene
Volume	128
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(93)90163-W
State	Published - Jun 15 1993

Keywords

Ecotin
fusion proteins
phage M13
phagemid
protein engineering
serine protease

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(93)90163-W

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title = "Trypsin display on the surface of bacteriophage",

abstract = "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.",

keywords = "Ecotin, fusion proteins, phage M13, phagemid, protein engineering, serine protease",

author = "{R. Corey}, David and Shiau, {Andrew K.} and Yang Qing and Janowski, {Bethany A.} and Craik, {Charles S.}",

note = "Funding Information: This authors are supported by NSF grants DMB-8904956an d EET-8807179(t o C.S.C.),a Damon Runyon-Walter Winchell Cancer Fund FellowshipD RG 1076( to D.R.C.), and a predoctoralf ellowshipf rom the Howard Hughes Medical Institute( to A.K.S.).",

year = "1993",

month = jun,

day = "15",

doi = "10.1016/0378-1119(93)90163-W",

language = "English (US)",

volume = "128",

pages = "129--134",

journal = "Gene",

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T1 - Trypsin display on the surface of bacteriophage

AU - R. Corey, David

AU - Shiau, Andrew K.

AU - Qing, Yang

AU - Janowski, Bethany A.

AU - Craik, Charles S.

N1 - Funding Information: This authors are supported by NSF grants DMB-8904956an d EET-8807179(t o C.S.C.),a Damon Runyon-Walter Winchell Cancer Fund FellowshipD RG 1076( to D.R.C.), and a predoctoralf ellowshipf rom the Howard Hughes Medical Institute( to A.K.S.).

PY - 1993/6/15

Y1 - 1993/6/15

N2 - 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.

AB - 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.

KW - Ecotin

KW - fusion proteins

KW - phage M13

KW - phagemid

KW - protein engineering

KW - serine protease

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DO - 10.1016/0378-1119(93)90163-W

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C2 - 8508953

AN - SCOPUS:0027231503

VL - 128

SP - 129

EP - 134

JO - Gene

JF - Gene

SN - 0378-1119

IS - 1

ER -

Trypsin display on the surface of bacteriophage

Abstract

Keywords

ASJC Scopus subject areas

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