Preliminary X-ray study of PapD Crystals from uropathogenic Escherichia coli

Holmgren, Anders; Brändén, Carl‐Ivar; Lindberg, Fredrik; Tennent, Jan M.

doi:10.1016/0022-2836(88)90109-x

Cited by 8 publications

(3 citation statements)

References 4 publications

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“…Such preparations will not only be valuable in the development of preventative therapies but will also provide material for X-ray crystallographic analyses of the P-pilus proteins. The structure of the PapD protein is interesting in itself; we expect that its features will soon be resolved, as the protein has recently been crystallized (17). Such approaches are essential if the intricacies of P-pilus biogenesis are to be understood.…”

Section: Discussionmentioning

confidence: 99%

PapD, a periplasmic transport protein in P-pilus biogenesis

et al. 1989

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The product of the papD gene of uropathogenic Escherichia coli is required for the biogenesis of digalactoside-binding P pili. Mutations within papD result in complete degradation of the major pilus subunit, PapA, and of the pilinlike proteins PapE and PapF and also cause partial breakdown of the PapG adhesin. The papD gene was sequenced, and the gene product was purified from the periplasm. The deduced amino acid sequence and the N-terminal sequence obtained from the purified protein revealed that PapD is a basic and hydrophilic peripheral protein. A periplasmic complex between PapD and PapE was purified from cells that overproduced and accumulated these proteins in the periplasm. Antibodies raised against this complex reacted with purified wild-type P pili but not with pili purified from a papE mutant. In contrast, anti-PapD serum did not react with purified pili or with the culture fluid of piliated cells. However, this serum was able to specifically precipitate the PapE protein from periplasmic extracts, confirming that PapD and PapE were associated as a complex. It is suggested that PapD functions in P-pilus biogenesis as a periplasmic transport protein. Probably PapD forms complexes with pilus subunits at the outer surface of the inner membrane and transports them in a stable configuration across the periplasmic space before delivering them to the site(s) of pilus polymerization.

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Section: Discussionmentioning

confidence: 99%

PapD, a periplasmic transport protein in P-pilus biogenesis

et al. 1989

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“…PapD was prepared as previously described (Holmgren et al, 1988). The peptide K(1Ј-19Ј) was prepared by F moc solid-phase synthesis and purified by reversed-phase HPLC (Karlsson et al, 1996).…”

Section: X-ray Crystallographymentioning

confidence: 99%

Periplasmic chaperone recognition motif of subunits mediates quaternary interactions in the pilus

Soto

Dodson

Ogg³

et al. 1998

The EMBO Journal

106

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The class of proteins collectively known as periplasmic immunoglobulin-like chaperones play an essential role in the assembly of a diverse set of adhesive organelles used by pathogenic strains of Gram-negative bacteria. Herein, we present a combination of genetic and structural data that sheds new light on chaperone-subunit and subunit-subunit interactions in the prototypical P pilus system, and provides new insights into how PapD controls pilus biogenesis. New crystallographic data of PapD with the C-terminal fragment of a subunit suggest a mechanism for how periplasmic chaperones mediate the extraction of pilus subunits from the inner membrane, a prerequisite step for subunit folding. In addition, the conserved N- and C-terminal regions of pilus subunits are shown to participate in the quaternary interactions of the mature pilus following their uncapping by the chaperone. By coupling the folding of subunit proteins to the capping of their nascent assembly surfaces, periplasmic chaperones are thereby able to protect pilus subunits from premature oligomerization until their delivery to the outer membrane assembly site.

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“…Elution of adhesin from galabiose-Sepharose with soluble receptor analogues yields soluble PapD-adhesin-receptor complexes that may be ideal for crystallization and analysis by x-ray crystallography. PapD crystals have already been obtained, so that three-dimensional structure can be determined (32). The ability of the soluble receptor analogues to elute adhesin from the receptor has also stimulated research in our laboratory to design compounds with a higher binding affinity for the adhesin than the naturally occurring receptor.…”

Section: I89mentioning

confidence: 99%

The PapG adhesin of uropathogenic Escherichia coli contains separate regions for receptor binding and for the incorporation into the pilus.

Hultgren

Lindberg

Magnusson

et al. 1989

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

172

122

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Most uropathogenic strains of Escherichia coli produce heteropolymeric organelles, known as P pili, that bind to the globoseries of glycolipids present in the urinary tract. The formation of a P pilus is the result of a family of related proteins being coordinately assembled into the structure in a defined order with the adhesin located exclusively at the tip. The preassembled digalactoside a-D-galactopyranosyl-(1--4)-.8-D-galactopyranose-binding adhesin was purified to homogeneity from the periplasmic space in a complex with the

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Preliminary X-ray study of PapD Crystals from uropathogenic Escherichia coli

Cited by 8 publications

References 4 publications

PapD, a periplasmic transport protein in P-pilus biogenesis

PapD, a periplasmic transport protein in P-pilus biogenesis

Periplasmic chaperone recognition motif of subunits mediates quaternary interactions in the pilus

The PapG adhesin of uropathogenic Escherichia coli contains separate regions for receptor binding and for the incorporation into the pilus.

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