Purification, inhibitory properties, amino acid sequence and identification of the reactive site of a new serine proteinase inhibitor from oil-rape (itBrassica napus) seed

Fabrizio Ceciliani, Fabrizio Bortolotti, Enea Menegatti, Severino Ronchi, Paolo Ascenzi, Sandro Palmieri

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

A new serine proteinase inhibitor, rapeseed trypsin inhibitor (RTI), has been isolated from rapeseed (Brassica napus var. oleifera) seed. The protein inhibits the catalytic activity of bovine β-trypsin and bovine α-chymotrypsin with apparent dissociation constants of 3.0 × 10-10 M and 4.1 × 10-7 M, at pH 8.0 and 21°C, respectively. The stoichiometry of both proteinase-inhibitor complexes is 1:1. The amino acid sequence of RTI consists of 60 amino acid residues, corresponding to an Mr, of about 6.7 kDa. The p1-pi, reactive site bond has been tentatively identified at position Arg20-Ile21. RTI shows no similarity to other serine proteinase inhibitors except the low molecular weight mustard trypsin inhibitor (MTI-2). RTI and MTI-2 could be members of a new class of plant serine proteinase inhibitors.

Original languageEnglish
Pages (from-to)221-224
Number of pages4
JournalFEBS Letters
Volume342
Issue number2
DOIs
Publication statusPublished - Apr 4 1994

Fingerprint

Brassica rapa
Serine Proteinase Inhibitors
Trypsin Inhibitors
Purification
Seed
Amino Acid Sequence
Catalytic Domain
Seeds
Oils
Amino Acids
Brassica napus
Chymotrypsin
Stoichiometry
Trypsin
Catalyst activity
Peptide Hydrolases
Molecular Weight
Molecular weight
Proteins

Keywords

  • Amino acid sequence
  • Brassica napus var. oleifera
  • Rapeseed
  • Serine proteinase inhibitor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification, inhibitory properties, amino acid sequence and identification of the reactive site of a new serine proteinase inhibitor from oil-rape (itBrassica napus) seed. / Ceciliani, Fabrizio; Bortolotti, Fabrizio; Menegatti, Enea; Ronchi, Severino; Ascenzi, Paolo; Palmieri, Sandro.

In: FEBS Letters, Vol. 342, No. 2, 04.04.1994, p. 221-224.

Research output: Contribution to journalArticle

Ceciliani, Fabrizio ; Bortolotti, Fabrizio ; Menegatti, Enea ; Ronchi, Severino ; Ascenzi, Paolo ; Palmieri, Sandro. / Purification, inhibitory properties, amino acid sequence and identification of the reactive site of a new serine proteinase inhibitor from oil-rape (itBrassica napus) seed. In: FEBS Letters. 1994 ; Vol. 342, No. 2. pp. 221-224.
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