Primary structure and reactive site of a novel wheat proteinase inhibitor of subtilisin and chymotrypsin.
The proteinase inhibitor WSCI, active in inhibiting bacterial subtilisin and a number of animal chymotrypsins, was purified from endosperm of exaploid wheat (Triticum aestivum, c.v. San Pastore) by ion exchange chromatography and its complete amino acid sequence was established by automated Edman degradation. WSCI consists of a single polypeptide chain of 72 amino acid residues, has a molecular mass of 8126.3 Da and a pl of 5.8. The inhibition constants (Ki) for Bacillus licheniformis subtilisin and bovine pancreatic alpha-chymotrypsin are 3.92 x 10(-9) M and 7.24 x 10(-9) M, respectively. The inhibitor contains one methionine and of tryptophan residue and has a high content of essential amino acids (41 over a total of 72 residues), but no cysteines. The primary structure of WSCI shows high similarity with barley subtilisin-chymotrypsin isoinhibitors of the Cl-2 type and with maize subtilisinchymotrypsin inhibitor MPI. Significant degrees of similarity were also found between sequences of WSCI and of other members of the potato inhibitor I family of the serine proteinase inhibitors. The wheat inhibitor WSCI has a single reactive site (the peptide bond between methionyl-48 and glutamyl-49 residues) as identified by affinity chromatography and sequence analysis.
Poerio
Elia
E
Dipartimento di Agrobiologia e Agrochimica, Università della Tuscia, I-01100 Viterbo, Italy.
Di Gennaro
Simone
S
Di Maro
Antimo
A
Farisei
Francesca
F
Ferranti
Pasquale
P
Parente
Augusto
A
D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000145:classification / Q000201:enzymology; D001665:Binding Sites; D002417:Cattle; D002846:Chromatography, Affinity; D002918:Chymotrypsin / Q000037:antagonists & inhibitors*; D008969:Molecular Sequence Data; D017421:Sequence Analysis / Q000379:methods; D017386:Sequence Homology, Amino Acid; D015842:Serine Proteinase Inhibitors / Q000235:genetics* / Q000302:isolation & purification; D020860:Subtilisin / Q000037:antagonists & inhibitors*; D014908:Triticum / Q000737:chemistry*
D015842:Serine Proteinase Inhibitors; D002918:Chymotrypsin; D020860:Subtilisin