The Structure of E. coli IgG-Binding Protein D Suggests a General Model for Bending and Binding in Trimeric Autotransporter Adhesins

Leo, Jack C.; Lyskowski, Andrzej Franciszek; Hattula, Katarina; Hartmann, M.D.; Schwarz, Heinz; Butcher, Sarah J.; Linke, Dirk; Lupas, Andrei N.; Goldman, Adrian

doi:10.1016/j.str.2011.03.021

Cited by 63 publications

(79 citation statements)

References 46 publications

(84 reference statements)

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“…For further fine tuning, the flexibility conferred by HANS may be delimited by FGG domains. The importance of flexibility for function has been pinpointed in detail for different TAAs, including the Moraxella catarrhalis adhesin UspA1 (26), Haemphilus influenzae Hia (21), and E. coli EibD (11). All of these adhesins bind to large receptors or matrix proteins on the surface of host cells, which disallows a rigid, straight fiber.…”

Section: Structure Determination Of Sada Fragments and Reconstruction Ofmentioning

confidence: 99%

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Complete fiber structures of complex trimeric autotransporter adhesins conserved in enterobacteria

Hartmann

Grin

Dunin-Horkawicz

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Trimeric autotransporter adhesins (TAAs) are modular, highly repetitive surface proteins that mediate adhesion to host cells in a broad range of Gram-negative pathogens. Although their sizes may differ by more than one order of magnitude, they all follow the same basic head-stalk-anchor architecture, where the head mediates adhesion and autoagglutination, the stalk projects the head from the bacterial surface, and the anchor provides the export function and attaches the adhesin to the bacterial outer membrane after export is complete. In complex adhesins, head and stalk domains may alternate several times before the anchor is reached. Despite extensive sequence divergence, the structures of TAA domains are highly constrained, due to the tight interleaving of their constituent polypeptide chains. We have therefore taken a "domain dictionary" approach to characterize representatives for each domain type by X-ray crystallography and use these structures to reconstruct complete TAA fibers. With SadA from Salmonella enterica, EhaG from enteropathogenic Escherichia coli (EHEC), and UpaG from uropathogenic E. coli (UPEC), we present three representative structures of a complex adhesin that occur in a conserved genomic context in Enterobacteria and is essential in the infection process of uropathogenic E. coli. Our work proves the applicability of the dictionary approach to understanding the structure of a class of proteins that are otherwise poorly tractable by high-resolution methods and provides a basis for the rapid and detailed annotation of newly identified TAAs.coiled coil | β-layer

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Section: Structure Determination Of Sada Fragments and Reconstruction Ofmentioning

confidence: 99%

“…It is followed by an extended and typically coiled-coil rich stalk, which projects the head from the bacterium and often provides binding sites for host serum factors (10,11). The protein ends in a membrane anchor (2).…”

mentioning

confidence: 99%

Complete fiber structures of complex trimeric autotransporter adhesins conserved in enterobacteria

Hartmann

Grin

Dunin-Horkawicz

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…For example, UspA1 showed a bending after binding to CAECAM-1 and fibronectin (Agnew et al, 2011). Similarly, the E. coli autotransporter EibD also bends from a saddle-like structure after binding to its ligand IgG (Leo et al, 2011). This kind of bending has only been observed after interaction with host ligands.…”

Section: Discussionmentioning

confidence: 99%

“…For instance, in some of the TAAs, interaction of ligands causes a bend in the trimer (Agnew et al, 2011;Leo et al, 2011). Unlike those proteins, the Hsf fibril is twisted after bending and the bended region may have loose or tight interactions with the first half part of the fibre.…”

Section: Discussionmentioning

confidence: 99%

Haemophilus influenzae surface fibril (Hsf) is a unique twisted hairpin-like trimeric autotransporter

Singh

Jubair

Mörgelin

et al. 2015

International Journal of Medical Microbiology

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The Haemophilus surface fibril (Hsf) is an extraordinary large (2413 amino acids) trimeric autotransporter, present in all encapsulated Haemophilus influenzae. It contributes to virulence by directly functioning as an adhesin. Furthermore, Hsf recruits the host factor vitronectin thereby inhibiting the host innate immune response resulting in enhanced survival in serum. Here we observed by electron microscopy that Hsf appears as an 100 nm long fibril at the bacterial surface albeit the length is approximately 200 nm according to a bioinformatics based model. To unveil this discrepancy, we denaturated Hsf at the surface of Hib by using guanidine hydrochloride (GuHCl). Partial denaturation induced in the presence of GuHCl unfolded the Hsf molecules, and resulted in an increased length of fibres in comparison to the native trimeric form. Importantly, our findings were also verified by E. coli expressing Hsf at its surface. In addition, a set of Hsf-specific peptide antibodies also indicated that the N-terminal of Hsf is located near the C-terminal at the base of the fibril. Taken together, our results demonstrated that Hsf is not a straight molecule but is folded and doubled over. This is the first report that provides the unique structural features of the trimeric autotransporter Hsf. Highlights Trimeric autotransporters are important virulence factors of Gram negative bacteria, and have a "lollipop-shape" structure or straight trimeric strand.  Haemophilus surface fibril is involved in adherence to host epithelium and for recruitment of vitronectin.  Here we show for the first time that Haemophilus surface fibril has a twisted hairpin-like structure.

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“…Interestingly, the EibD head and stalk structure revealed the stalk to coil in a left-handed direction from the β-barrel structure, and then switch to a right-handed-coil at approximately halfway (Leo et al, 2011). Crystal structures have also been solved for partial domains of other Gram-negative trimeric AT proteins, including the Yersinia adhesion YadA (Nummelin et al, 2004, Alvarez et al, 2010, Hia (Meng et al, 2006b, Yeo et al, 2004, UspA1 (Conners et al, 2008), SadA (Hartmann et al, 2009), BadA (Szczesny et al, 2008) and BpA (Edwards et al, 2010).…”

Section: Type Vc: Trimeric At Proteinsmentioning

confidence: 99%

Autotransporter proteins of Uropathogenic Escherichia coli: regulation, characterisation and the role of periplasmic proteins in their expression

Nichols¹

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Uropathogenic Escherichia coli (UPEC) are the primary cause for all urinary tract infections.Autotransporter (AT) proteins are virulence factors secreted by the type V secretion pathway. This project examined the prevalence, regulation and expression of the vacuolating AT toxin (Vat), and the role of periplasmic chaperone proteins in AT translocation.The vacuolating autotransporter (AT) toxin (Vat) contributes to Uropathogenic Escherichia coli (UPEC) fitness during systemic infection. Vat is a serine protease AT of Enterobacteriaceae (SPATE) with cytotoxic activity. Here we characterised Vat and investigated its regulation in UPEC.We assessed the prevalence of vat in a collection of 45 UPEC urosepsis strains and showed it that was present in 31 (68%) of the isolates. The isolates containing the vat gene corresponded to three major E. coli sequence types (ST12, 73 and 95) and these strains secreted the Vat protein.Further analysis of the vat genetic locus identified a conserved gene located directly downstream of vat that encodes a putative MarR-like transcriptional regulator, which we termed vatX. The vatvatX genes were present in the UPEC reference strain CFT073 and RT-PCR revealed both genes are co-transcribed. Over-expression of vatX in CFT073 led to a 3-fold increase in vat gene transcription. The vat promoter region contained three putative nucleation sites for the global transcriptional regulator H-NS; thus the hns gene was mutated in CFT073 (to generate CFT073hns).Western blot analysis using a Vat-specific antibody revealed a significant increase in Vat expression in CFT073hns compared to wild-type CFT073. Direct H-NS binding to the vat promoter region was demonstrated using purified H-NS in combination with electrophoresis mobility shift assays. Finally, Vat-specific antibodies were detected in plasma samples from urosepsis patients iv Declaration by authorThis thesis is composed of my original work, and contains no material previously published or written by another person except where due reference has been made in the text. I have clearly stated the contribution by others to jointly-authored works that I have included in my thesis.

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The Structure of E. coli IgG-Binding Protein D Suggests a General Model for Bending and Binding in Trimeric Autotransporter Adhesins

Cited by 63 publications

References 46 publications

Complete fiber structures of complex trimeric autotransporter adhesins conserved in enterobacteria

Complete fiber structures of complex trimeric autotransporter adhesins conserved in enterobacteria

Haemophilus influenzae surface fibril (Hsf) is a unique twisted hairpin-like trimeric autotransporter

Autotransporter proteins of Uropathogenic Escherichia coli: regulation, characterisation and the role of periplasmic proteins in their expression

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