title: The effect of variation within inhibitory domains on the activity of pea protease inhibitors from the Bowman-Birk class.
abstract: We have investigated the properties of variant pea seed protease inhibitors, homologous to the anti-carcinogenic Bowman-Birk inhibitor (BBI) from soybean but differing most significantly in amino acid sequences at the two independent sites of protease inhibition. The pea protease inhibitors were expressed, using Aspergillus niger, with yields of up to 23 mg secreted recombinant protein per litre of media. The recombinant proteins showed protease inhibitory activity and were deduced to be disulphide-bonded correctly; limited post-translational processing had occurred at the amino-terminal ends of all proteins. Differences in trypsin and chymotrypsin specific inhibitory activities, and in inhibition constants, were observed in studies of the two recombinant variants and BBI.
authors.affiliation: Department of Metabolic Biology, John Innes Centre, Norwich Research Park, NR4 7UH, UK. alfonso.clemente@sainsbury-laboratory.ac.uk
authors.forename: Alfonso
authors.initials: A
authors.lastname: Clemente
authors.forename: Donald A
authors.initials: DA
authors.lastname: MacKenzie
authors.forename: David J
authors.initials: DJ
authors.lastname: Jeenes
authors.forename: Claire
authors.initials: C
authors.lastname: Domoney
mesh_terms: D000595:Amino Acid Sequence; D001203:Ascomycota / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000201:enzymology* / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D011110:Polymorphism, Genetic; D017434:Protein Structure, Tertiary / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014357:Trypsin / Q000737:chemistry; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000737:chemistry; D014361:Trypsin Inhibitors / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics
chemical_list: D010940:Plant Proteins; D011994:Recombinant Proteins; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D014361:Trypsin Inhibitors; D002918:Chymotrypsin; D014357:Trypsin