Isolation and characterization of cytoplasmic fibrils from treponemes.
Electron microscopy of Triton X-100-treated whole cells of an oral treponeme, Treponema sp. strain E-21, revealed that six cytoplasmic fibrils (CFs) helically wound as a bundle in the cytoplasm. The CFs were isolated and purified by disruption and solubilization of the cells followed by CsCl density gradient centrifugation. The purified CF preparation contained mostly fibrils of about 9 nm in width and very small amounts of thinner strands of about 3 nm in diameter. The CFs were apparently seen to be a tubular structure, but the isolated CFs had narrowed sites of about 4-5 nm in width lacking lumen-like images, possibly representing twisted sites. Thus, the CF did not seem to be a tubular structure. The purified CFs were composed of one major 82 kDa protein and a few minor proteins. The CFs were destructed by treatment with proteases, 8 M urea or 4 M guanidine hydrochloride. Very low tyrosine content (0.76 mol %) and lack of methionine were characteristic features for the 82 kDa protein. The CF preparations from the other five treponemes including Treponema phagedenis and T. denticola also had 82 kDa proteins as a major component, and the 82 kDa proteins of all of the treponemes had a common antigen when examined by using antiserum against the 82 kDa protein from Treponema sp. strain E-21. Furthermore, the 82 kDa protein was demonstrated to be a principal component of the CFs of all the treponemes by immunoelectron microscopy.
Masuda
K
K
Department of Food Microbiology, Tokushima University School of Medicine.
Kawata
T
T
D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000302:isolation & purification; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D008854:Microscopy, Electron; D008868:Microtubule Proteins / Q000302:isolation & purification; D008870:Microtubules / Q000648:ultrastructure*; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols; D014208:Treponema / Q000648:ultrastructure*; D014461:Ultracentrifugation
D000596:Amino Acids; D001426:Bacterial Proteins; D003902:Detergents; D008868:Microtubule Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol