G. Layhschmitt et R. Herrmann, SPATIAL ARRANGEMENT OF GENE-PRODUCTS OF THE P1 OPERON IN THE MEMBRANEOF MYCOPLASMA-PNEUMONIAE, Infection and immunity, 62(3), 1994, pp. 974-979
The spatial arrangement of three P1 operon-encoded proteins-attachment
protein P1 (ORF5 gene product) and the ORF6-derived proteins of 40 an
d 90 kDa-in the membrane of Mycoplasma pneumoniae M129 was investigate
d by nearest-neighbor analysis. For these studies, the homobivalent, t
hiol-cleavable, and nonmembrane-permeating cross-linking reagent 3,3'-
dithiobis(sulfosuccinimidylpropionate) (DTSSP) was used. The cross-lin
ked proteins were isolated by immunoprecipitation with antibodies dire
cted against fusion proteins of selected regions of the 40-kDa, the 90
-kDa, or the P1 protein. The individual components of the immunoprecip
itated protein complexes were identified by sodium dodecyl sulfate-pol
yacrylamide gel electrophoresis, autoradiography, and immunoblot analy
sis. This study showed that the P1 protein, the ORF6 gene product, and
an unidentified 30-kDa protein were linked to each other in the intac
t bacterial membrane by the reagent DTSSP, indicating that these prote
ins are located at a maximal distance of 12 A Angstrom (1A Angstrom= 0
.1 nm) on the tip structure of M. pneumoniae.