Molecular characterization of the structural gene for the lactoferrin receptor of the meningococcal strain H44/76.
The meningococcal lactoferrin receptor is a promising vaccine candidate since it seems to be antigenically rather stable. Monoclonal antibodies against this protein reacted with more than 50% of the strains tested. To gain further insight in its variability, the IbpA gene from strain H44/76, encoding this protein, was cloned and sequenced. This strain does not cross-react with monoclonal antibodies that recognize LbpA of strain BNCV. The deduced amino acid sequence was found to be 95% homologous to the previously established sequence of LbpA of strain BNCV. A topology model was proposed for LbpA, making use of the sequence comparisons and some general rules for the folding of outer membrane proteins. The protein is supposed to traverse the membrane 26 times in a beta-sheet conformation. The epitope recognized by the monoclonals was mapped and found to reside in the largest predicted surface-exposed loop. No iron-regulation of LbpA expression was found in E. coli, probably because IbpA is located in an operon, the promoter of which was not cloned. Upstream of IbpA, a part of an open reading frame was found. Whereas the LbpA protein shows homology to the transferrin-binding protein 1 (Tbp1), the putative protein encoded by the open reading frame upstream of IbpA shows extensive homology to Tbp2, suggesting that iron-acquisition from lactoferrin, like from transferrin, requires two specific proteins in the outer membrane. The upstream open reading frame is tentatively designated IbpB.
Pettersson
A
A
Department of Molecular Cell Biology, Utrecht University, The Netherlands.
Klarenbeek
V
V
van Deurzen
J
J
Poolman
J T
JT
Tommassen
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D004926:Escherichia coli / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D007501:Iron / Q000378:metabolism; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D009876:Operon / Q000235:genetics; D011487:Protein Conformation; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA
D011956:Receptors, Cell Surface; C038340:lactoferrin receptors; D007501:Iron