Ordered translocation of 987P fimbrial subunits through the outer membrane of Escherichia coli

Cao, Jiangbei; Khan, Abdul Salam; Bayer, Manfred; Schifferli, Dieter M.

doi:10.1128/jb.177.13.3704-3713.1995

Cited by 27 publications

(41 citation statements)

References 51 publications

(48 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The inserted DNA was sequenced to confirm PCR accuracy. The recombinant proteins were expressed by induction with isopropyl-␤-D-thiogalactopyranoside (IPTG) in E. coli BL21(DE3) and purified by metal chelation chromatography, as described previously (34). The resulting proteins did not have their signal sequences and contained 6-histidine tags at their C-terminal ends.…”

Section: Methodsmentioning

confidence: 99%

Adhesive Properties of YapV and Paralogous Autotransporter Proteins of Yersinia pestis

et al. 2015

View full text Add to dashboard Cite

Section: Methodsmentioning

confidence: 99%

Adhesive Properties of YapV and Paralogous Autotransporter Proteins of Yersinia pestis

et al. 2015

View full text Add to dashboard Cite

“…Shifts in the sizes of the MalE fusion proteins indicated that the fusion proteins were produced and exported to the periplasm. These fusion proteins were purified with amylose columns, and the histidine-tagged FasE protein was purified with a nickel column as described previously (3). Rabbits were immunized with the purified proteins (three rabbits per antigen) as described before (3).…”

Section: Methodsmentioning

confidence: 99%

“…1 and Table 1). These fimbriae consist of a helical repeat of the major subunit, FasA, with FasG and FasF located both at the tip of the fimbriae and at intervals along the fimbriae as minor subunits (3,12). We have previously shown that FasG is the 987P adhesin and recognizes a glycoprotein on piglet intestinal brush borders.…”

mentioning

confidence: 99%

“…We have previously shown that FasG is the 987P adhesin and recognizes a glycoprotein on piglet intestinal brush borders. FasF was proposed to be inserted between FasG and FasA during fimbrial biogenesis (3,17). The usher protein, located in the outer membrane, directs translocation of the subunits from the periplasm during fimbrial biogenesis, and we have previously shown that FasD is the usher for 987P fimbriae (30).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Identification of major and minor chaperone proteins involved in the export of 987P fimbriae

1996

Self Cite

View full text Add to dashboard Cite

The 987P fimbriae of Escherichia coli consist mainly of the major subunit, FasA, and two minor subunits, FasF and FasG. In addition to the previously characterized outer membrane or usher protein FasD, the FasB, FasC, and FasE proteins are required for fimbriation. To better understand the roles of these minor proteins, their genes were sequenced and the predicted polypeptides were shown to be most similar to periplasmic chaperone proteins of fimbrial systems. Western blot (immunoblot) analysis and immunoprecipitation of various fas mutants with specific antibody probes identified both the subcellular localizations and associations of these minor components. FasB was shown to be a periplasmic chaperone for the major fimbrial subunit, FasA. A novel periplasmic chaperone, FasC, which stabilizes and specifically interacts with the adhesin, FasG, was identified. FasE, a chaperone-like protein, is also located in the periplasm and is required for optimal export of FasG and possibly other subunits. The use of different chaperone proteins for various 987P subunits is a novel observation for fimbrial biogenesis in bacteria. Whether other fimbrial systems use a similar tactic remains to be discovered.There are many ways in which diarrheogenic enteric bacteria both recognize and attach to specific host receptors and ensure direct delivery of enterotoxins (5) or signal transduction molecules (16) to intestinal epithelial cells. Enterotoxigenic Escherichia coli utilize fimbriae which optimize the attachment of this pathogen by bridging the bacteria and the host's enterocytes.We have studied the 987P fimbriae of enterotoxigenic E. coli, which mediate attachment to piglet intestines. The eight genes (fasA to fasH) which encode all the proteins required for fimbriation have been localized to a plasmid (31), and functions have been identified for five of their products ( Fig. 1 and Table 1). These fimbriae consist of a helical repeat of the major subunit, FasA, with FasG and FasF located both at the tip of the fimbriae and at intervals along the fimbriae as minor subunits (3, 12). We have previously shown that FasG is the 987P adhesin and recognizes a glycoprotein on piglet intestinal brush borders. FasF was proposed to be inserted between FasG and FasA during fimbrial biogenesis (3, 17). The usher protein, located in the outer membrane, directs translocation of the subunits from the periplasm during fimbrial biogenesis, and we have previously shown that FasD is the usher for 987P fimbriae (30). It has been demonstrated by others that FasH (FapR) is a transcriptional activator required for the expression of fasA and possibly other fas genes (19).A number of additional proteins, apart from the exported subunits, are required for the production of functional fimbriae. Fimbrial subunits are exported by a Sec-dependent mechanism from the cytoplasm to the periplasm and eventually through the outer membrane. After crossing the cytoplasmic membrane, the subunits are thought to be in a quasinative conformation and are able to aggregate...

show abstract

“…A fourth construct, pZS468, was prepared by inverse PCR (41) using pZS669 as template and two primers designed to delete residues Lys-30 to Asn-109 from the full-length histone H1 (Table I). The histone H1 protein and protein fragments were expressed in E. coli BL21(DE3) and analyzed by SDS-PAGE as described previously (26,34).…”

Section: Bacterial Strains and Plasmidmentioning

confidence: 99%

Histone H1 Proteins Act As Receptors for the 987P Fimbriae of Enterotoxigenic Escherichia coli

Zhu¹,

Chen

Choi³

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

The tip adhesin FasG of the 987P fimbriae of enterotoxigenic Escherichia coli mediates two distinct adhesive interactions with brush border molecules of the intestinal epithelial cells of neonatal piglets. First, FasG attaches strongly to sulfatide with hydroxylated fatty acyl chains. This interaction involves lysine 117 and other lysine residues of FasG. Second, FasG recognizes specific intestinal brush border proteins that migrate on a sodium-dodecyl sulfate-polyacrylamide gel like a distinct set of 32-35-kDa proteins, as shown by ligand blotting assays. The protein sequence of high performance liquid chromatography-purified tryptic fragments of the major protein band matched sequences of human and murine histone H1 proteins. Porcine histone H1 proteins isolated from piglet intestinal epithelial cells demonstrated the same SDS-PAGE migration pattern and 987P binding properties as the 987P-specific protein receptors from porcine intestinal brush borders. Binding was dose-dependent and shown to be specific in adhesion inhibition and gel migration shift assays. Moreover, mapping of the histone H1 binding domain suggested that it is located in their lysine-rich C-terminal domains. Histone H1 molecules were visualized on the microvilli of intestinal epithelial cells by immunohistochemistry and electron microscopy. Taken together these results indicated that the intestinal protein receptors for 987P are histone H1 proteins. It is suggested that histones are released into the intestinal lumen by the high turnover of the intestinal epithelium. Their strong cationic properties can explain their association with the negatively charged brush border surfaces. There, the histone H1 molecules stabilize the sulfatide-fimbriae interaction by simultaneously binding to the membrane and to 987P. Enterotoxigenic Escherichia coli (ETEC)1 cause diarrhea in mammals by expressing at least one type of enteroadhesive fimbria and one type of enterotoxin. By adhering to intestinal epithelial cells, localized multiplication of an ETEC strain can progress to mucosal surface colonization and concomitant effective enterotoxin delivery. This was illustrated with hostspecific ETEC strains in both animals and human volunteers (1-5). ETEC are the most important etiologic agents of both neonatal and postweaning diarrhea in pigs (6, 7). 987P-fimbriated ETEC cause diarrhea in neonatal piglets in the United States (8), Europe (9, 10), Asia (11-15), and Central and South America (16 -18). In addition to this widespread distribution, the recent identification of human ETEC strains with 987P-like fimbriae (19, 20) on various continents highlights the evolutionary adaptability of these fimbriae (21).The 987P fimbria consists of the helical arrangement of protein subunits along a filamentous axis (22,23). It is a heteropolymeric structure that is made up of one major subunit, FasA, and two minor subunits, FasF and FasG (24). Fimbriae are not produced in the absence of any of these subunits (25). Electron microscopy and export studies indicated that FasG ...

show abstract

Ordered translocation of 987P fimbrial subunits through the outer membrane of Escherichia coli

Cited by 27 publications

References 51 publications

Adhesive Properties of YapV and Paralogous Autotransporter Proteins of Yersinia pestis

Adhesive Properties of YapV and Paralogous Autotransporter Proteins of Yersinia pestis

Identification of major and minor chaperone proteins involved in the export of 987P fimbriae

Histone H1 Proteins Act As Receptors for the 987P Fimbriae of Enterotoxigenic Escherichia coli

Contact Info

Product

Resources

About