Bovine pancreatic trypsin inhibitor-trypsin complex as a detection system for recombinant proteins

J. Borjigin, J. Nathans

Research output: Contribution to journalArticle

Abstract

Bovine pancreatic trypsin inhibitor (BPTI) binds to trypsin and anhydrotrypsin (an enzymatically inactive derivative of trypsin) with affinities of 6 x 10-14 and 1.1 x 10-13 M, respectively. We have taken advantage of the high affinity and specificity of this binding reaction to develop a protein tagging system in which biotinylated trypsin or biotinylated anhydrotrypsin is used as the reagent to detect recombinant fusion proteins into which BPTI has been inserted. Two proteins, opsin and growth hormone, were used as targets for insertional mutagenesis with BPTI. In each case, both domains of the fusion protein appear to be correctly folded. The fusion proteins can be specifically and efficiently detected by biotinylated trypsin or biotinylated anhydrotrypsin, as demonstrated by staining of transfected cells, protein blotting, affinity purification, and a mobility shift assay in SDS/polyacrylamide gels.

Original languageEnglish (US)
Pages (from-to)337-341
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number1
DOIs
StatePublished - Jan 12 1993

Keywords

  • human growth hormone
  • protein tagging
  • rhodopsin

ASJC Scopus subject areas

  • General

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