title: Cholera toxin is synthesized in precursor form on free polysomes in Vibrio cholerae 569B.
abstract: Membrane-bound and free polysomes have been isolated from Vibrio cholerae 569B. Nacent polypeptide chains were completed in a cell-free translation mixture containing Escherichia coli S-300 extracts and [3H]leucine or [35S]methionine. Cholera toxin-related polypeptides synthesized in vitro were immunologically detected after treatment with either anti-subunit A or anti-subunit B serum. Immunoreactive translation products were removed from reaction mixtures with formalinized Cowan's strain of Staphylococcus aureus, electrophoresed on sodium dodecyl sulfate-polyacrylamide gels, and visualized by fluorography. Anti-subunit A serum precipitated two major polypeptide species (molecular weights 52,000 and 45,000) from translation mixtures programed with free polysomes, whereas anti-subunit B serum precipitated only the 45,000-molecular-weight polypeptide. No cholera toxin-related polypeptides were detectable in translation mixtures programed with membrane-bound polysomes. Purified subunit A and cholera toxin competed for anti-subunit A binding sites and blocked the immunoprecipitation of the 35S-labeled 52,000- and 45,000-dalton polypeptides from in vitro translation mixtures. The data presented suggest that cholera toxin is synthesized in the cytoplasm in a precursor form on free polysomes and is secreted post-translationally.
mesh_terms: D002772:Cholera Toxin / Q000096:biosynthesis*; D008970:Molecular Weight; D011132:Polyribosomes / Q000378:metabolism*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000096:biosynthesis*; D014734:Vibrio cholerae / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D011498:Protein Precursors; D002772:Cholera Toxin