Ordered translocation of 987P fimbrial subunits through the outer membrane of Escherichia coli.
The 987P fimbria of enterotoxigenic Escherichia coli is a heteropolymeric structure which consists essentially of a major FasA subunit and a minor subunit, the FasG adhesin. The latter harbors the binding moiety for receptor molecules on piglet intestinal epithelial cells. In this study, anti-FasF antibody probes were developed and used to demonstrate that the FasF protein represents a new minor fimbrial component. FasF was identified in highly purified fimbriae, and its sequence demonstrated significant levels of similarity with that of FasA. Immune electron microscopy localized both the FasG and FasF proteins at the fimbrial tip as well as at broken ends and at various intervals along the fimbrial length. The presence of these minor proteins in purified 987P fimbriae was corroborated by enzyme-linked immunosorbent assay inhibitions. Finally, the use of nonfimbriated fasG, fasF, and fasA mutants indicated that subunit translocation through the outer membrane follows a specific order, FasG being the first, FasF being the second, and FasA being the third type of exported subunit. Since fimbriae are thought to grow from the base, FasG is proposed to be a tip adhesin and FasF is proposed to be a linker molecule between the adhesin and the fimbrial shaft. Moreover, export of FasG (or FasF) in the absence of FasF (or FasA) indicates that during the process of fimbrial biogenesis in the outer membrane, translocating events precede the initiation of subunit heteropolymerization.
Cao
J
J
Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia 19104, USA.
Khan
A S
AS
Bayer
M E
ME
Schifferli
D M
DM
D018830:Adhesins, Escherichia coli / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000302:isolation & purification / Q000378:metabolism; D000954:Antigens, Surface / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D008954:Models, Biological; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013997:Time Factors
D018830:Adhesins, Escherichia coli; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001426:Bacterial Proteins; C035391:antigen 987P, E coli; D036781:Fimbriae Proteins