Structural Differences between Trypsin Inhibitors Isolated from Cucurbitaceae Family Seeds: Immunological, NMR and CD Studies

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Abstract
By manifold immunizations of rabbits with virgin or modified trypsin inhibitor III from squash seeds and trypsin inhibitor II b from cucumber seeds, specific antibodies were produced. In double immunodiffusion the anti-squash inhibitor antibody also gave weak precipitate arcs with inhibitor I from squash, inhibitor II from summer squash and with inhibitor I from zucchini, but not with inhibtor II b from cucumber seeds. The genus of Cucurbita trypsin inhibitors, preincubated with the antibody, lost their antitrypsin activity. The antibody showed a significantly weaker effect on the activity of the inhibitor from cucumber seeds. 1H-NMR and CD [circular dichroism] spectra also confirm structural differences between typsin inhibitors from the genus of Cucurbita and the cucumber (Cucumis) inhibitor.