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PMID: 1548058 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cloning, expression, and DNA sequence analysis of genes encoding nontypeable Haemophilus influenzae high-molecular-weight surface-exposed proteins related to filamentous hemagglutinin of Bordetella pertussis.

Infection and immunity ·Vol. 60 ·No. 4 ·1992-04-00 ·Pages 1302-13

Barenkamp SJ, Leininger E

Abstract

A group of high-molecular-weight surface-exposed proteins of nontypeable Haemophilus influenzae are major targets of human serum antibody (S. J. Barenkamp and F. F. Bodor, Pediatr. Infect. Dis. J. 9:333-337, 1990). To further characterize these proteins, we cloned and sequenced genes encoding two related high-molecular-weight proteins from a prototype nontypeable Haemophilus strain. The gene encoding a 120-kDa Haemophilus protein consisted of a 4.4-kbp open reading frame, and the gene encoding a 125-kDa protein consisted of a 4.6-kbp open reading frame. The first 1,259 bp of the two genes were identical. Thereafter, the sequences began to diverge, but overall they were 80% identical, and the derived amino acid sequences showed 70% identity. A protein sequence homology search demonstrated similarity between the derived amino acid sequences of both cloned genes and the derived amino acid sequence of the gene encoding filamentous hemagglutinin, a surface protein produced by the gram-negative pathogen Bordetella pertussis. Antiserum raised against a recombinant protein encoded by the 4.6-kbp open reading frame recognized both the 120- and the 125-kDa proteins in the prototype strain as well as antigenically related high-molecular-weight proteins in 75% of a collection of 125 epidemiologically unrelated nontypeable H. influenzae strains. The antiserum directed against the recombinant protein also recognized purified filamentous hemagglutinin. A murine monoclonal antibody to filamentous hemagglutinin recognized both the 120-kDa and the 125-kDa protein in the prototype strain as well as proteins identical to those recognized by the recombinant-protein antiserum in 35% of the nontypeable H. influenzae strain collection. Thus, we have identified and partially characterized a group of highly immunogenic surface-exposed proteins of nontypeable H. influenzae which are related to the filamentous hemagglutinin of B. pertussis.

MeSH Terms
Adhesins, Bacterial Amino Acid Sequence Antibodies, Monoclonal Bacterial Proteins/genetics Base Sequence Blotting, Western Bordetella pertussis/genetics Cloning, Molecular Enzyme-Linked Immunosorbent Assay Gene Expression Regulation, Bacterial Genes, Bacterial Haemophilus influenzae/genetics Hemagglutinins/genetics Membrane Proteins/genetics Molecular Sequence Data Plasmids Restriction Mapping Sequence Homology, Nucleic Acid Virulence Factors, Bordetella
Chemicals
Adhesins, Bacterial Antibodies, Monoclonal Bacterial Proteins Hemagglutinins Membrane Proteins Virulence Factors, Bordetella filamentous hemagglutinin adhesin, Bordetella pertussis
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Barenkamp S J
Edward Mallinckrodt Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri.
Leininger E
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.60.4.1302-1313.1992
Published
1992-04-00
Pages
1302-13
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC256997
Subset
IM
Grants
NIAID NIH HHS · AI21707 · United States
Databases
GENBANK
M84615, M84616
External Links
PubMed source DOI: 10.1128/iai.60.4.1302-1313.1992 PMC Full Text View journal details
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