A chimeric mini-trypsin inhibitor derived from the oil rape proteinase inihibitor type III

Maurizio Trovato, Bruno Maras, Fabio Polticelli, Paolo Costantino, Paolo Ascenzi

Research output: Contribution to journalArticlepeer-review


The design of chimeric proteins is a major field of interest in structural biology and biotechnology. The successful design of the chimeric protein composed by the minimized reactive site domain of the low-molecular-mass trypsin inhibitor from Brassica napus (var. oleifera) seed (Ser3-Lys35; mini-RTI-III) and murine dihydrofolate reductase (DHFR) is reported here. The DHFR-mini-RTI-III chimeric protein was expressed in Escherichia coli, purified by metal-chelate affinity chromatography and oxidatively refolded. The affinity of the purified and refolded DHFR-mini-RTI-III for bovine trypsin (K = 5.0 x 10-10 M) was closely similar to that determined for native RTI-III (K = 2.9 x 10-10 M), at pH 8.2 and 22.0°C. DHFR-mini-RTI-III may be regarded as a tool in structure-function studies and for developing multifunctional and multidomain proteinase inhibitors. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)817-820
Number of pages4
JournalBiochemical and Biophysical Research Communications
Issue number3
Publication statusPublished - Sep 7 2000


  • Chimeric oil rape mini-trypsin-inhibitor
  • E. coli expression
  • Inhibitory properties
  • Oil rape (Brassica napus var. oleifera) trypsin inhibitor
  • Oxidative refolding
  • White mustard (Sinapis alba L.) trypsin inhibitor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology


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