Molecular characterization of LbpB, the second lactoferrin-binding protein of Neisseria meningitidis

Mol Microbiol. 1998 Feb;27(3):599-610. doi: 10.1046/j.1365-2958.1998.00707.x.

Abstract

The lbpA gene of Neisseria meningitidis encodes an outer membrane lactoferrin-binding protein and shows homology to the transferrin-binding protein, TbpA. Previously, we have detected part of an open reading frame upstream of lbpA. The putative product of this open reading frame, tentatively designated lbpB, showed homology to the transferrin-binding protein TbpB, suggesting that the lactoferrrin receptor, like the transferrin receptor, consists of two proteins. The complete nucleotide sequence of lbpB was determined. The gene encodes a 77.5 kDa protein, probably a lipoprotein, with homology, 33% identity to the TbpB of N. meningitidis. A unique feature of LbpB is the presence of two stretches of negatively charged residues, which might be involved in lactoferrin binding. Antisera were raised against synthetic peptides corresponding to the C-terminal part of the putative protein and used to demonstrate that the gene is indeed expressed. Consistent with the presence of a putative Fur binding site upstream of the lbpB gene, expression of both LbpA and LbpB was proved to be iron regulated in Western blot experiments. The LbpB protein appeared to be less stable than TbpB in SDS-containing sample buffer. Isogenic mutants lacking either LbpA or LbpB exhibited a reduced ability to bind lactoferrin. In contrast to the lbpB mutant, the lbpA mutant was completely unable to use lactoferrin as a sole source of iron.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Outer Membrane Proteins*
  • Bacterial Proteins*
  • Base Sequence
  • Carrier Proteins / chemistry*
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism
  • Cloning, Molecular
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli
  • Genes, Bacterial
  • Immunoblotting
  • Lactoferrin / metabolism*
  • Molecular Sequence Data
  • Mutagenesis
  • Neisseria meningitidis / chemistry
  • Neisseria meningitidis / genetics*
  • Neisseria meningitidis / growth & development
  • Neisseria meningitidis / metabolism
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • Porins / metabolism
  • Receptors, Cell Surface*
  • Restriction Mapping
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid

Substances

  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • Carrier Proteins
  • DNA-Binding Proteins
  • Porins
  • Receptors, Cell Surface
  • lactoferrin binding proteins, bacterial
  • Lactoferrin

Associated data

  • GENBANK/AF022781