From the inside out--processing of the Chlamydial autotransporter PmpD and its role in bacterial adhesion and activation of human host cells.
Polymorphic membrane protein (Pmp)21 otherwise known as PmpD is the longest of 21 Pmps expressed by Chlamydophila pneumoniae. Recent bioinformatical analyses annotated PmpD as belonging to a family of exported Gram-negative bacterial proteins designated autotransporters. This prediction, however, was never experimentally supported, nor was the function of PmpD known. Here, using 1D and 2D PAGE we demonstrate that PmpD is processed into two parts, N-terminal (N-pmpD), middle (M-pmpD) and presumably third, C-terminal part (C-pmpD). Based on localization of the external part on the outer membrane as shown by immunofluorescence, immuno-electron microscopy and immunoblotting combined with trypsinization, we demonstrate that N-pmpD translocates to the surface of bacteria where it non-covalently binds other components of the outer membrane. We propose that N-pmpD functions as an adhesin, as antibodies raised against N-pmpD blocked chlamydial infectivity in the epithelial cells. In addition, recombinant N-pmpD activated human monocytes in vitro by upregulating their metabolic activity and by stimulating IL-8 release in a dose-dependent manner. These results demonstrate that N-PmpD is an autotransporter component of chlamydial outer membrane, important for bacterial invasion and host inflammation.
Wehrl
Wolfgang
W
Max-Planck Institute for Infection Biology, Department of Molecular Biology, Schumannstr. 21/22, 10117 Berlin, Germany.
Brinkmann
Volker
V
Jungblut
Peter R
PR
Meyer
Thomas F
TF
Szczepek
Agnes J
AJ
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D002462:Cell Membrane / Q000648:ultrastructure; D002689:Chlamydia / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007822:Laryngeal Neoplasms; D008969:Molecular Sequence Data; D009000:Monocytes / Q000382:microbiology; D010446:Peptide Fragments / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D014357:Trypsin; D014407:Tumor Cells, Cultured
D001425:Bacterial Outer Membrane Proteins; D010446:Peptide Fragments; C484024:polymorphic membrane protein 21, Chlamydophila pneumoniae; D014357:Trypsin