Abstract
The complete amino acid sequence of a white kidney bean (Phaseolus vulgaris) α-amylase inhibitor (PHA-I), which is composed of two kinds of glycopolypeptide subunits, α and β, was established by conventional methods. The polypeptide molecular weight of PHA-I determined by the light-scattering technique, considered together with the sequence molecular weights revealed for the subunits, indicated that PHA-I has the subunit stoichiometry of (αβ)2 complex. Inhibition test of PHA-I with increasing amounts of porcine pancreatic α-amylase (PPA) suggested that an inactive 2:1 complex is formed between PPA and PHA-I. In fact, two complexes differing from each other in the molar ratio of PPA to PHA-I were separated by gel filtration, and molecular weight estimation by the light-scattering technique confirmed that they are complexes of PHA-I with one or two PPA molecules. The binding of PPA to PHA-I appeared to follow simple binomial statistics, suggesting that two binding sites on PHA-I are independent and of high affinity for PPA.
