Spatial arrangement of gene products of the P1 operon in the membrane of Mycoplasma pneumoniae.
The spatial arrangement of three P1 operon-encoded proteins--attachment protein P1 (ORF5 gene product) and the ORF6-derived proteins of 40 and 90 kDa--in the membrane of Mycoplasma pneumoniae M129 was investigated by nearest-neighbor analysis. For these studies, the homobivalent, thiol-cleavable, and nonmembrane-permeating cross-linking reagent 3,3'-dithiobis(sulfosuccinimidylpropionate) (DTSSP) was used. The cross-linked proteins were isolated by immunoprecipitation with antibodies directed against fusion proteins of selected regions of the 40-kDa, the 90-kDa, or the P1 protein. The individual components of the immunoprecipitated protein complexes were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, autoradiography, and immunoblot analysis. This study showed that the P1 protein, the ORF6 gene product, and an unidentified 30-kDa protein were linked to each other in the intact bacterial membrane by the reagent DTSSP, indicating that these proteins are located at a maximal distance of 12 A (1 A = 0.1 nm) on the tip structure of M. pneumoniae.
Layh-Schmitt
G
G
ZMBH, Mikrobiologie, Universität Heidelberg, Federal Republic of Germany.
Herrmann
R
R
D018829:Adhesins, Bacterial; D001345:Autoradiography; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D009177:Mycoplasma pneumoniae / Q000737:chemistry* / Q000235:genetics; D009876:Operon
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; C035471:adhesin, Mycoplasma pneumoniae