Isolation and characterization of a wound-induced trypsin inhibitor from alfalfa leaves

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Abstract
A trypsin inhibitor from leaves of field-grown alfalfa plants was purified; it is the same trypsin inhibitor that is wound-induced in leaves of young growth chamber grown plants. This inhibitor accounts for the major trypsin inhibitory activity found in both, field-grown and wound-induced plants. The inhibitor exhibits a MW of .apprx. 7500 and is specific for trypsin with a Ki of 1 .times. 10-10 M. Analysis of the purified inhibitor by cation-exchange high-performance liquid chromatography revealed the presence of 4 isoinhibitor species that have identical immunological and inhibitory properties. The amino acid analysis of the 4 spp. indicates small but significant differences. Immunological double diffusion comparisons of the alfalfa inhibitor with the Bowman-Birk and Kunitz soybean inhibitors did not reveal any cross-reactivity although the amino acid content of the alfalfa inhibitor resembles those of Bowman-Birk family members.

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