The role of the P2' position of Bowman-Birk proteinase inhibitor in the inhibition of trypsin. Studies on P2' variation in cyclic peptides encompassing the reactive site loop.
The role of the P2' residue in proteinase inhibitors of the Bowman-Birk family was investigated using synthetic cyclic peptides based on the reactive site loop of the inhibitor. A series of 21 variants having different P2' residues was tested for inhibition of trypsin, and the rate at which they were hydrolysed by this enzyme was also measured. Variation at P2' was found to result in marked differences in inhibitory potency, with the best sequence (Ile) having a Ki value of 9 nM. Peptides with P2' Gly, Pro or Glu failed to demonstrate any measurable inhibition (Ki>1 mM). The peptides also displayed significant differences in the rates at which they were hydrolysed, which varied by over three orders of magnitude between the difference sequences. There was found to be overall correlation between the Ki value and the rate of hydrolysis, with peptides that inhibited best also being hydrolysed more slowly. The results are discussed in light of the sequence information for Bowman-Birk inhibitor proteins.
Gariani
T
T
Department of Chemistry, Imperial College of Science, Technology and Medicine, South Kensington, London SW7 2AY, UK.
McBride
J D
JD
Leatherbarrow
R J
RJ
D001665:Binding Sites / Q000235:genetics; D006868:Hydrolysis; D007700:Kinetics; D010456:Peptides, Cyclic / Q000138:chemical synthesis* / Q000235:genetics; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000737:chemistry*
D010456:Peptides, Cyclic; D014358:Trypsin Inhibitor, Bowman-Birk Soybean