Amino acid sequence of a Bowman-Birk proteinase inhibitor from pea seeds

Abstract

Trypsin inhibitors from winter pea seeds (c.v. Frilene) have been purified by ammonium sulfate precipitation, gel filtration, and anion and cation exchange chromatography and shown to consist of six protease inhibitors (PSTI I, II, III, IVa, IVb, and V). Their molecular weights were determined by electrospray mass spectrometry as 6916, 6807, 7676, 7944, 7848 and 7844 D, respectively, and the sequences of the first 20 N-terminal amino acid residues of these six inhibitors were found to be identical. The complete amino acid sequence of PSTI IVa was determined. This protein comprises a total of 72 residues and has 14 cysteines, all involved in disulfide bridges. Comparison of the sequence of PSTI IVa with those of other leguminous Bowman-Birk type inhibitors revealed that PSTI could be classified as a group III inhibitor, closely related to Vicia faba and Vicia angustifolia inhibitors.

Proteinase Inhibitors from Pea Seeds: Purification and Characterization

“…According to their amino acid composition as well as their molecular masses, these six pea trypsin inhibitors may belong to the Bowman-Birk family. It was further confirmed by the similarity of their N-terminal sequences with those of both V. faba and V. angustifolia inhibitors as well as with that of PSTI IVa, which was recently sequenced by ourselves (Ferrasson et al, 1995). In particular, the six inhibitors possess the Pro-Pro sequence at positions 19-20 and the Lys16-Ser17 reactive peptide bond.…”

“…The presence of various Bowman-Birk isoinhibitors in most of the species studied (Odany and Ikenaka, 1977;Norioka and Ikenaka, 1983;Ishikawa et al, 1985) is attributed either to the expression by distinct genes and/or to the posttranslational proteolytic cleavage of few amino acids at the N-or C-terminal end of the inhibitors. In the case of peas, the existence of many isoforms has also been observed (Weder and Hory, 1972;Valdebouze et al, 1980;Tome ´et al, 1981;Gaborit et al, 1989;Domoney et al, 1993;Ferrasson et al, 1995); some of them have been characterized. It was shown by cDNA sequencing (Domoney et al, 1993) as well as protein sequencing (Ferrasson et al, 1995) that some of these inhibitors belonged to the Bowman-Birk family.…”

“…In the case of peas, the existence of many isoforms has also been observed (Weder and Hory, 1972;Valdebouze et al, 1980;Tome ´et al, 1981;Gaborit et al, 1989;Domoney et al, 1993;Ferrasson et al, 1995); some of them have been characterized. It was shown by cDNA sequencing (Domoney et al, 1993) as well as protein sequencing (Ferrasson et al, 1995) that some of these inhibitors belonged to the Bowman-Birk family. Complementary information is needed to know if the other isoforms were also members of this family.…”

Complete Amino Acid Sequence of the Lentil Trypsin−Chymotrypsin Inhibitor LCI-1.7 and a Discussion of Atypical Binding Sites of Bowman−Birk Inhibitors

“…Because one of the porcine chymotrypsin components, chymotrypsin C, cleaves peptide bonds also after glutamine and methionine (26), and a chymotrypsinogen similar to chymotrypsinogen C from porcine pancreas has been isolated from human pancreatic juice (27), binding of HCT at Gln18 close to that reactive site has Comparison of the primary structures of group III Bowman−Birk inhibitors from legume seeds: LCI-1.7, trypsin−chymotrypsin inhibitor from lentils (this paper); LCI-L1 F , trypsin−chymotrypsin inhibitor fragment from L. culinaris leaves (10), included for comparison (sequence from nucleic acid; residues 1−42 not shown, probably propeptide and/or signal as in garden pea); PSTI IVA and IVB, trypsin inhibitors from winter pea (P. sativum cv. Frilene) (11,12); PSI TI-5−72, trypsin inhibitor from garden pea (P. sativum cv. Birte; sequence from cDNA) (13); PSI TI-F, trypsin−chymotrypsin inhibitor from garden pea (P. sativum cv.…”

10.5937/ratpov51-5072 = Significance of genetic resources of cool season annual legumes: II: Neglected and underutilised crops

“…The local landraces of fenugreek may be a source of the tolerance to crenate broomrape, a significant parasitic plant in grain legumes in the Mediterranean (Fernández-Aparicio et al 2008). Good results were achieved if fenugreek is intercropped with grain legumes, such as faba bean (Fernández-Aparicio et al 2011). On the other hand, there is a recent report on the susceptibility of the local Tunisian landraces of fenugreek to fetid broomrape, although to an extremely small extent (Amri et al 2009).…”