Endo-1,4-beta-glucanase in the cell wall of stems of auxin-treated pea seedlings.
Endo-1,4-beta-glucanase induced by treatment of pea seedlings with 2,4-D was extracted from a preparation of the walls of epicotyl cells. The beta-glucanase was purified by chromatography on DEAE-cellulose, affinity chromatography on Con A-Sepharose and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The activity of beta-glucanase was retained after removal of SDS and extraction from polyacrylamide gels. The band of a protein (46 kDa), that corresponded to the activity of endo-1,4-beta-glucanase, was injected directly into mice for preparation of antiserum and the protein was also subjected to amino acid sequencing after blotting onto a membrane. Western blot analysis showed that the antiserum obtained bound to a 46-kDa polypeptide and recognized endo-1,4-beta-glucanase. The N-terminal sequence of the 46-kDa polypeptide revealed some homology to abscission endo-1,4-beta-glucanases of bean and avocado fruit.
Hayashi
T
T
Basic Research Laboratory, Ajinomoto Co. Inc., Kawasaki, Japan.
Ohsumi
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D002473:Cell Wall / Q000201:enzymology*; D002480:Cellulase / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D004790:Enzyme Induction; D007887:Fabaceae / Q000196:embryology / Q000201:enzymology* / Q000648:ultrastructure; D007210:Indoleacetic Acids / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D010946:Plants, Medicinal; D012639:Seeds / Q000201:enzymology / Q000648:ultrastructure
D000906:Antibodies; D007210:Indoleacetic Acids; D002480:Cellulase