Biochemical and structural analyses of microtubules in the pellicular membrane of Leishmania tropica.
The structure of the major protein of the pellicular membrane of Leishmania tropica was investigated. This protein is composed of two polypeptides, of ca. 50,000 d molecular weight, that were found to cross-react immunologically with the alpha and beta subunits of pig brain tubulin. The polypeptides and pig brain tubulin subunits were partially digested with S. aureus V8 protease, and the peptides obtained analysis by SDS-polyacrylamide gel electrophoresis. A comparison of the patterns showed that the beta subunits of Leishmania and pig tubulin have very similar primary structures, while the alpha subunits have evolved divergently. These experiments demonstrate that the major polypeptides found in the pellicular membrane of L. tropica are alpha and beta subunits of tubulin. Immunoelectron microscopy indicates that the tubulin is located in the microtubules associated with the pellicular membrane of Leishmania. Arrays of microtubules were prepared by nonionic detergent treatment of the cells and observed by electron microscopy after negative staining. Optical diffraction reveals a 5 nm spacing between protofilaments in the microtubule and a 4 nm axial periodicity corresponding to the tubulin subunits. The pitch of the shallow left-hand three-start helix is 12 degrees. A distance of 47 nm separates each microtubule from the next. These data show that the dimensions and supramolecular organization of the tubulin subunits in the microtubules are identical in the pellicular membrane of L. tropica and in mammalian brain.
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D007891:Leishmania / Q000032:analysis / Q000648:ultrastructure*; D046911:Macromolecular Substances; D008566:Membranes / Q000032:analysis; D008870:Microtubules / Q000032:analysis* / Q000648:ultrastructure; D010455:Peptides / Q000032:analysis; D013552:Swine; D014404:Tubulin / Q000032:analysis* / Q000276:immunology
D046911:Macromolecular Substances; D010455:Peptides; D014404:Tubulin