An immunological assessment of lysosomal enzymes and other macromolecules sulfated during vegetative growth of Dictyostelium discoideum

Davis SJ, Wheldrake JF, Freeze HH. (1988), J Cell Biochem. 38, 113-6

Abstract

Western blotting and immunoprecipitation data indicated that lysosomal enzymes represent a subset of the sulfated macromolecules present in vegetative Dictyostelium discoideum amoebae and account for less than 2.5% of the total sulfate incorporated during vegetative growth. These data suggest that the majority of the highly sulfated macromolecules of vegetative D. discoideum amoebae are not related to the lysosomal enzymes.

Key figure: Western blot analysis of the sulfated macromolecules

After electrophoresis on 12% polyacrylamide gels in the presence of sodium dodecyl sulfate, [³⁵S]sulfate-labelled macromolecules from vegetative D. discoideum strain NP73 amoebae were visualized by fluorography (lane 1). The same macromolcules separated in another lane of the same gel were transferred to nitrocellulose for an incubation with mLE1 antibody. Macromolecules recognized by the antibody were visualized with a second antibody conjugated to alkaline phosphatase (lane 2).