Endo-1,4-{beta}-Glucanase in the Cell Wall of Stems of Auxin-Treated Pea Seedlings

Endo-1,4-ß-glucanase induced by treatment of pea seedlingswith 2,4-D was extracted from a preparation of the walls ofepicotyl cells. The ß-glucanase was purified by chromatographyon DEAE-cellulose, affinity chromatography on Con A-Sepharoseand SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The activityof ß-glucanase was retained after removal of SDS andextraction from polyacrylamide gels. The band of a protein (46kDa), that corresponded to the activity of endo-1,4-ß-glucanase,was injected directly into mice for preparation of antiserumand the protein was also subjected to amino acid sequencingafter blotting onto a membrane. Western blot analysis showedthat the antiserum obtained bound to a 46-kDa polypeptide andrecognized endo-1,4-ß-glucanase. The N-terminal sequenceof the 46-kDa polypeptide revealed some homology to abscissionendo-1,4-ß-glucanases of bean and avocado fruit. (Received September 29, 1993; Accepted January 20, 1994)