An amphipathic sulphated glycoconjugate of Leishmania: characterization with monoclonal antibodies.

摘要: A major glycoconjugate of Leishmania tropica identified by two monoclonal antibodies was shown to be an externally oriented, amphipathic membrane antigen shed into the culture medium in which parasites grow. This molecule could labelled metabolically with [3H]glucose, [3H]galactose, [32P]phosphate and [35S]sulphate. It migrated as a polydisperse band upon electrophoresis SDS-polyacrylamide gels, spanning region gel corresponding apparent mol. wt. 20 000-67 000 daltons. An apparently identical family molecules on surface living promastigotes using galactose oxidase [3H]-sodium borohydride. released supernatant over period several hours. Detection 3H- or 35S-labelled required days exposure autoradiographs, but novel blotting technique nitrocellulose coated antibody allowed rapid detection charge shift electrophoresis, Western dot blotting. The electrophoretic mobility agarose relative its Triton X-100 increased presence deoxycholate, decreased cetyl trimethyl-ammonium bromide, indicating properties consistent insertion lipid bilayer membrane. Using dot-blotting detected all virulent avirulent clones LRC-L137 additional isolates L. (LRC-L287 LRC-L251), not donovani mexicana, previously described specificity antibodies. However, general approaches used this paper showed that (LRC-L52) mexicana (LRC-L94) synthesize similar, antigenically distinct glycoconjugate.