(14-38, 30-51) Double-disulphide intermediate in folding of bovine pancreatic trypsin inhibitor: A two-dimensional 1H nuclear magnetic resonance study

Carlo P.M. van Mierlo*, Nigel J. Darby, David Neuhaus, Thomas E. Creighton

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

79 Citations (Scopus)

Abstract

An analogue of the BPTI folding intermediate that contains only the disulphide bonds between Cys14 and Cys38 and between Cys30 and Cys51 has been prepared in Escherichia coli by protein engineering methods. The other two Cys residues of native BPTI (at positions 5 and 55) have been replaced by Ser. Essentially complete proton resonance assignments of the analogue were obtained by employing two-dimensional 1H nuclear magnetic resonance techniques. The intermediate has a more extended conformation in the N-terminal (residues 1 to 7) region and there are other differences in the C-terminal (residues 55 to 58) region. The remainder of the protein is substantially identical to native BPTI. The conformational properties of the analogue can explain several aspects of the kinetic role that the normal (14-38, 30-51) intermediate plays in the folding of BPTI.

Original languageEnglish
Pages (from-to)353-371
JournalJournal of Molecular Biology
Volume222
Issue number2
DOIs
Publication statusPublished - 20 Nov 1991

Keywords

  • bovine pancreatic trypsin inhibitor (BPTI)
  • disulphide bonds
  • folding intermediate
  • n.m.r.
  • protein folding

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