Purification and characterization of Haemophilus influenzae pili, and their structural and serological relatedness to Escherichia coli P and mannose-sensitive pili.

Guerina, Nicholas G.; Langermann, Solomon; Schoolnik, Gary K.; Keßler, Tobias; Goldmann, Donald A.

doi:10.1084/jem.161.1.145

Cited by 41 publications

(44 citation statements)

References 19 publications

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“…This observation is not surprising because NH2-terminal sequence homologies have been reported in type 1, P, and H. influenzae fimbriae (12,14). Although there appears to be no homology in the NH2-terminal sequence between 987P and the type 1 fimbrial subunit (21), common …”

Section: Resultsmentioning

confidence: 52%

Assembly of a chemically synthesized peptide of Escherichia coli type 1 fimbriae into fimbria-like antigenic structures

Abraham¹,

Beachey²

1987

J Bacteriol

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Escherichia coli type 1 fimbriae are composed of subunits, each of which comprises 158 amino acids. We synthesized a copy of a 13-residue peptide, located near the NH2 terminus of the fimbrial subunit, that assumed some of the properties of type 1 fimbriae. At pH 5.5 the synthetic peptide autoassembled into fibrillar structures that resembled type 1 fimbriae except that they appeared less rigid and rodlike. A quaternary structure-specific monoclonal antibody against type 1 fimbriae recognized the synthetic peptide in the assembled but not the unassembled state. Furthermore, when the synthetic peptide was injected in its fimbrial conformation into rabbits, it evoked antibodies that reacted with type 1 fimbriae isolated from E. coli.Type 1 fimbriae on the surface of various members of the family Enterobacteriaceae have been shown to mediate the attachment of these microorganisms to mannose-containing receptors on eucaryotic cells (1,2,7,9,20,23,24). Purified type 1 fimbriae agglutinate guinea pig erythrocytes and bind to mucosal epithelial cells in a D-mannose-sensitive manner. Brinton (7) has shown that type 1 fimbriae are composed of 17-kilodalton protein subunits that are assembled into a right-handed helix that is 7 nm in diameter. Each turn of the helix consists of 3 1/8 subunits. The subunits are held together by hydrophobic bonds which are highly resistant to disruption but can be dissociated by boiling in acid (7) or treating with saturated solutions of guanidine hydrochloride (11). The dissociated subunits have been shown to reassemble in vitro into fimbrial structures (2, 11). This property is not unique to type 1 fimbriae; the fimbriae of Pseudomonas aeruginosa (27) and 987P of Escherichia coli (24a) have been shown to have similar properties.The mechanism of fimbrial autoassembly is not clear, although results of studies of the primary structure of several different types of fimbriae have demonstrated a high degree of homology in their NH2-terminal amino acid sequences (12-16, 21, 22, 26). Because of these homologies, we decided to synthesize several overlapping peptides of the first 35 residues of the NH2-terminal region of E. coli type 1 fimbriae. Here we report that one of these peptides, containing residues 23 to 35, assembles itself into fibrillar structures resembling native type 1 fimbriae. We show that antibodies raised against the assembled synthetic peptide react both with fimbrial subunits and assembled type 1 fimbriae isolated from E. coli. Moreover, monoclonal antibodies directed against quaternary structural determinants of native type 1 fimbriae react with the assembled, but not the unassembled, synthetic fimbriae.MATERIALS AND METHODS Synthesis of peptides. Peptides were synthesized by solidphase techniques (19) and purified as described previously (6). The purity of the peptide product was 'determined by high-performance liquid chromratography on a reverse phase column and by quantitative amino acid analyses (6).

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Section: Resultsmentioning

confidence: 52%

Assembly of a chemically synthesized peptide of Escherichia coli type 1 fimbriae into fimbria-like antigenic structures

Abraham¹,

Beachey²

1987

J Bacteriol

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“…The strong correlation between pili and S motility observed by Kaiser (15) suggests a function in cohesion, because the role of pili as attachment organelles has been demonstrated in a variety of pathogenic bacteria (2,9,17,21,23). M. xanthus pili about one cell length long were observed in a tuft at one of the cell poles (15).…”

Section: Discussionmentioning

confidence: 97%

Cell surface properties correlated with cohesion in Myxococcus xanthus

1988

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The gliding behavior of Myxococcus xanthus cells is controlled by two multigene systems, A and S, which encode information for adventurous and social behaviors, respectively. The S system can be genetically disrupted through mutation, such as a dsp mutation, or phenotypically disrupted by treating cells with the diazo dye Congo red (Arnold and Shimkets, J. Bacteriol. 170:5765-5770, 1988). One of the functions controlled by the S system is cell agglutination. Immediately after the induction of agglutination, wild-type cells begin to form aggregates, and within 30 min the cells are packed side-to-side in clumps containing thousands of cells. Changes in the cohesive properties of S+ cells are correlated with changes in the topology of the cell surface observed by electron microscopy. Two types of cell-associated appendages were observed on wild-type cells: thin filaments (ca. 5 nm in diameter), which have been called fimbriae or pili, at one cell pole, and thick, flaccid ifiaments (ca. 50 nm in diameter), referred to as fibrils, at both the sides and tips of cells. Cohesion was correlated with the secretion of the thick fibrils, which coat the cell surface and form an extracellular matrix in which the cells are interconnected. Several lines of evidence suggest that these thick fibrils are involved in cohesion. First, Dsp cells were unable to agglutinate or secrete this extracellular material. Second, wild-type cells which were treated with Congo red neither agglutinated nor secreted the extracellular fibrils. Finally, removal of the Congo red from wild-type cells restored cohesion and also restored production of the thick fibrils. Attempts to estimate the efficiency with which two cells cohered following collision suggested that under optimal conditions, one in three collisions resulted in stable contact. The collision efficiency decreased linearly as the cell density increased, suggesting a cell density-dependent regulation of cohesion. Some aspects of gliding behavior can be explained in terms of an inducer and an inhibitor of S motility.

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“…Other virulence factors include lipopolysaccharide (Kaplan et al, 1988), IgA proteases (Farley et al, 1986), and fimbriae (Guerina et al, 1985). However, little information is available concerning the mechanism(s) or role of iron sequestration in the pathogenesis of Haemophilus infections.…”

Section: Introductionmentioning

confidence: 99%

Siderophore-independent acquisition of transferrin-bound iron by Haemophilus influenzae type b

Morton

Williams²

1990

Journal of General Microbiology

123

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The specificity by which Huemophilus species acquired iron from transferrin (TF) was investigated. In a plate bioassay H. injYuenzse used iron bound to human, bovine and rabbit TFs but not mouse, rat, dog, horse, guinea-pig, pig or ovo-TFs or human and bovine lactoferrins. In contrast, H. pleuropneumoniae used iron only from pig TF whilst H. parainfienzae was unable to utilize iron bound to any of the human or animal TFs tested. The inhibition of growth imposed on H. injhenzae type b strain Eagan by the addition of the synthetic iron chelator EDDA to the culture medium was reversed by 30% iron-saturated human TF added directly to the medium but not when the TF was contained inside a dialysis bag. Dot-blotting of whole cells revealed that human TF bound to the surface of bacteria cultured in iron-restricted but not in iron-plentiful media. Incubation of whole bacterial cells in the presence of the proteolytic enzyme trypsin also abolished TF-binding activity, suggesting that the TF receptor was a protein. In competition dot blotting experiments, human and bovine but not rabbit, dog, mouse or guinea-pig TFs blocked the binding of a horseradish peroxid-human TF conjugate. SDS-PAGE and Western blotting of outer membranes revealed the presence of a TF-binding protein of approximately 72 kDa. These results suggest that the acquisition of TF-bound iron by H. injYuenzae type b probably involves a direct interaction with an outer-membrane protein which shows some TF-species specificity.

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Purification and characterization of Haemophilus influenzae pili, and their structural and serological relatedness to Escherichia coli P and mannose-sensitive pili.

Cited by 41 publications

References 19 publications

Assembly of a chemically synthesized peptide of Escherichia coli type 1 fimbriae into fimbria-like antigenic structures

Assembly of a chemically synthesized peptide of Escherichia coli type 1 fimbriae into fimbria-like antigenic structures

Cell surface properties correlated with cohesion in Myxococcus xanthus

Siderophore-independent acquisition of transferrin-bound iron by Haemophilus influenzae type b

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