Identification of a genetic locus of Haemophilus influenzae type b necessary for the binding and utilization of heme bound to human hemopexin

Mark S. Hanson, Sharon E. Pelzel, J. O. Latimer, Ursula Muller-Eberhard, Eric J. Hansen

Research output: Contribution to journalArticle

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Abstract

The mechanism(s) used by Haemophilus influenzae to acquire the essential nutrient heme from its human host has not been elucidated. The heme carried by the high-affinity serum protein hemopexin is one potential source of this micronutrient in vivo. A colony-blot assay revealed that hemehuman hemopexin-binding activity was shared among most capsular serotype b strains of H. influenzae but was uncommon among other strains. We have identified a recombinant clone binding heme-human hemopexin from a H. influenzae type b (Hib) genomic library expressed in Escherichia coli. Both the Hib strain and the heme-hemopexin-binding clone expressed a polypeptide of ≈100 kDa that bound radiolabeled heme-hemopexin. Oligonucleotide linker insertion mutagenesis of the plasm id DNA from this recombinant clone was used to confirm that expression of the 100-kDa protein correlated with the heme-hemopexin-binding activity. Exchange of one of these mutant alleles into the Hib chromosome eliminated expression of both the 100-kDa protein and the heme-hemopexin-binding activity. Furthermore, this Hib mutant was unable to utilize heme-human hemopexin as a heme source. (.

Original languageEnglish (US)
Pages (from-to)1973-1977
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number5
StatePublished - 1992

Fingerprint

Hemopexin
Haemophilus influenzae type b
Genetic Loci
Heme
Clone Cells
Recombinant DNA
Genomic Library
Micronutrients
Insertional Mutagenesis
Haemophilus influenzae
Oligonucleotides
Blood Proteins
Chromosomes
Alleles
Escherichia coli
Food
Peptides

Keywords

  • Bacterial pathogenesis
  • Heme acquisition

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Identification of a genetic locus of Haemophilus influenzae type b necessary for the binding and utilization of heme bound to human hemopexin. / Hanson, Mark S.; Pelzel, Sharon E.; Latimer, J. O.; Muller-Eberhard, Ursula; Hansen, Eric J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 5, 1992, p. 1973-1977.

Research output: Contribution to journalArticle

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AU - Muller-Eberhard, Ursula

AU - Hansen, Eric J.

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