Molecular characterization and outer membrane association of a Chlamydia trachomatis protein related to the hsp70 family of proteins.
One route by which Chlamydia trachomatis is internalized into host endometrial epithelial cells is receptor-mediated endocytosis. Although this implies an adhesin-receptor interaction exists, specific chlamydial surface molecules have not been identified. We are investigating potential adhesin molecules using an in vitro functional assay to select for chlamydial recombinant Escherichia coli expressing an adherent phenotype. We have previously shown that E. coli JM109(pPBW58) attaches to epithelial cells by a specific process paralleling C. trachomatis and expresses at least three plasmid-encoded proteins (18, 28, and 82 kDa; Schmiel, D. H., Knight, S. T., Raulston, J. E., Choong, J., Davis, C. H., and Wyrick, P. B. (1991) Infect. Immun. 59, 4001-4012). In this report, we demonstrate that (i) the 82-kDa protein is associated with the outer membrane of both E. coli JM109-(pPBW58) and C. trachomatis serovar E elementary bodies; (ii) the plasmid-encoded protein is identical to the native chlamydial protein by mass, charge, antigenicity, and partial proteolytic peptide profiles; (iii) a highly homologous protein is present in C. trachomatis biovariant lymphogranuloma venereum; (iv) the 82-kDa protein is not covalently linked by disulfide bonds to other protein species in either E. coli JM109(pPBW58) or C. trachomatis; (v) sequence analysis of the open reading frame indicates this protein is a relative of the heat shock 70 family of proteins; and (vi) the inferred amino acid sequence contains a contiguous 73-amino acid region having 51% identity with the extracellular sperm receptor binding domain in Strongylocentrosus purpuratus (Foltz, K. R., Partin, J. S., and Lennarz, W. J. (1993) Science 259, 1421-1425). The potential involvement of an hsp70 protein in attachment may provide new insight on adherence mechanisms by obligate intracellular pathogens.
Raulston
J E
JE
Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill 27599-7290.
Davis
C H
CH
Schmiel
D H
DH
Morgan
M W
MW
Wyrick
P B
PB
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D001483:Base Sequence; D002692:Chlamydia trachomatis / Q000737:chemistry*; D003001:Cloning, Molecular; D004220:Disulfides / Q000737:chemistry; D006360:Heat-Shock Proteins / Q000737:chemistry*; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D011956:Receptors, Cell Surface / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012617:Sea Urchins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004220:Disulfides; D006360:Heat-Shock Proteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C035027:egg surface sperm receptor