Cloning and characterization of Neisseria meningitidis genes encoding the transferrin-binding proteins Tbp1 and Tbp2

Legrain, M; Mazarin, V.; Irwin, Scan W.; Bouchon, Bernadette; Quentin‐Millet, Marie‐José; Jacobs, Eric J.; Schryvers, Anthony B.

doi:10.1016/0378-1119(93)90348-7

Cited by 144 publications

(171 citation statements)

References 30 publications

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“…The TF receptor is composed of the 100 kDa transmembrane protein, Tbp1, and the 85 kDa lipoprotein, Tbp2, encoded by the genes tbpA and tbpB, respectively (Schryvers and Morris, 1988a;Cornelissen et al, 1992;Legrain et al, 1993;Anderson et al, 1994;Cornelissen and Sparling, 1994;Cornelissen and Sparling, 1996). The LF receptor is composed of the 103 kDa transmembrane protein Lbp1 and a lipoprotein, Lbp2, analogous to Tbp2 encoded by the genes lbpA and lbpB, respectively (Schryvers and Morris, 1988b;Pettersson et al, 1993;1994a,b;Biswas and Sparling, 1995;Bonnah et al, 1995;Lewis et al, 1996;our unpublished observations).…”

Section: Introductionmentioning

confidence: 99%

Cloning and functional characterization of Neisseria gonorrhoeae tonB, exbB and exbD genes

Biswas

Anderson

Sparling

1997

Molecular Microbiology

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SummaryNeisseria gonorrhoeae is able to utilize iron (Fe) from a variety of sources including transferrin (TF) and lactoferrin (LF). To gain insight into the molecular mechanisms used by gonococci to scavenge Fe from TF and LF, we cloned a 3.5 kb segment of wild-type DNA that repaired the defect in tlu mutants, which are unable to take up Fe from either TF or LF despite exhibiting apparently normal ligand binding to the receptor. Nucleotide sequence determination identified three open reading frames (ORFs), designated ORF1, ORF2, and ORF3, which were arranged in tandem. The deduced amino acid sequence of the 852 bp ORF1 encoded a 28 kDa protein that exhibited 26-32% identity with TonB proteins of nine other bacteria. The 663 bp ORF2 predicted a 24 kDa protein and the 435 bp long ORF3 predicted a 15 kDa protein. These predicted protein sequences exhibited 32-38% and 24-36% identity, respectively, with ExbB and ExbD proteins of three other bacteria. Thus, the sequence comparison identified the ORF1, ORF2 and ORF3 as gonococcal homologues of the E. coli tonB, exbB and exbD genes. An insertional mutation in the tonB homologue resulted in the failure of gonococci to grow with TF, LF or human haemoglobin (HB) as sole Fe sources and in the inability to take up 55 Fe from TF and LF. The tonB mutation did not prevent the utilization of Fe from citrate (CT) or haemin (HM). Binding of TF, LF and HB to whole cells in a solid-phase binding assay was largely unaffected by the tonB mutation. We conclude that the pathways for utilization of Fe bound to TF, LF and HB but not to HM or CT were dependent on the TonB system.

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Section: Introductionmentioning

confidence: 99%

Cloning and functional characterization of Neisseria gonorrhoeae tonB, exbB and exbD genes

Biswas

Anderson

Sparling

1997

Molecular Microbiology

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“…, 1995). Tbp2 is also accessible at the surface and appears to be a lipoprotein presumably anchored in the membrane by its lipid tail Legrain et al, 1993). In Neisseria meningitidis, Tbp2 appears to be required for iron acquisition from transferrin (Irwin e t al., 1993), whereas in Neisseria gonorrboeae and Haemophilus influenxae it may be important but not essential for this process (Gray-Owen et al, 1995;Anderson e t al., 1994).…”

Section: Introductionmentioning

confidence: 99%

Sequence, genetic analysis, and expression of Actinobacillus pleuropneumoniae transferrin receptor genes

Gonzalez

Rosteck

et al. 1995

Microbiology

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The tbpA and *pi? genes encoding the transferrin receptor proteins T b p l and Tbp2 from a serotype 7 strain of Actinobacillus pleuropneumoniae were cloned, sequenced, and expressed in Escherichia coli. The tbpB gene was preceded by putative promoter and regulatory sequences and was separated from the downstream tbpA gene by a 13 bp intercistronic sequence suggesting that the two genes may be coordinately transcribed. Determination of the nucleotide sequence of this region facilitated PCR amplification of the tbp region from a serotype 1 strain for comparative purposes. The deduced amino acid sequences of the T b p l proteins had regions of homology with Neisseria Lbp and Tbpls and with TonB-dependent outer membrane (OM) receptors of E. coli. The deduced amino acid sequences of the Tbp2 proteins were nearly identical to those presented in previous studies. Upon high-level expression of the tbpA gene, a large proportion of the recombinant T b p l was found in inclusion bodies and could not be aff inity-isolated with immobilized porcine transferrin. Most of the remaining expressed T b p l was present in the OM fraction, was expressed at the surface of E. coli cells, and retained binding activity that was specific for the Globe of porcine transferrin. Although recombinant Tbp2 was found in inclusion bodies during high-level expression, a significant proportion was associated with a novel OM fraction that appeared in sucrose density gradients which was distinct from the OM fraction containing recombinant Tbpl. The recombinant Tbp2 was accessible at the surface yet was unable to bind porcine transferrin. In contrast to previous observations, the binding by recombinant Tbp2 was specific for the Globe of porcine transferrin. These results indicate that the A. pleuropneumoniae transferrin receptor proteins have similar properties to the receptor proteins in Neisseria spp. and Haemophilus influenzae, and that functional studies performed with recombinant receptor proteins need to consider differences in processing and export of these proteins when expressed in heterologous hosts.

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“…The genes encoding Tbpl and Thp2 have been isolated from two N. meningitidis group B strains, B16B6 and M982 (12), which were chosen for the differences observed in electrophoretic mobilities and antigenic properties of their Tbp2s (16). They were shown to be organized in tandem, tbp2 being followed by tbpl, with an 87-bp intergenic region.…”

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“…Tbp2 is produced as a precursor containing an N-terminal 20-amino-acid (aa) signal peptide whose cleavage follows a lipidation step on the first amino acid (cysteine) of the mature protein (lla, 12). Tbpl is produced as a precursor with a conventional secretion signal (12). Isogenic mutations were recently obtained in N. meningitidis, and bcth Tbpl and Tbp2 were shown to be able to bind hTf at the bacterial surface and to be essential for growth on hTf as an iron source (10).…”

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confidence: 99%

Characterization of a highly structured domain in Tbp2 from Neisseria meningitidis involved in binding to human transferrin

et al. 1994

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The binding of iron-loaded human transferrin at the surface of Neisseria meningitidis is mediated by two polypeptides, Tbp1 and Tbp2. Predicted Tbp amino acid sequences from N. meningitidis strains are highly divergent. This variability is particularly pronounced throughout the Tbp2 polypeptide. In this study, a highly structured and extremely stable Tbp2 domain of about 270 to 290 amino acids which is involved in the binding to transferrin and whose position is well conserved has been characterized. The conservation of such a remarkable structure in a very divergent protein domain (there is only 43% amino acid identity within this region) suggests that is plays an essential biological role and raises a number of questions regarding tbp2 evolution.

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Cloning and characterization of Neisseria meningitidis genes encoding the transferrin-binding proteins Tbp1 and Tbp2

Cited by 144 publications

References 30 publications

Cloning and functional characterization of Neisseria gonorrhoeae tonB, exbB and exbD genes

Cloning and functional characterization of Neisseria gonorrhoeae tonB, exbB and exbD genes

Sequence, genetic analysis, and expression of Actinobacillus pleuropneumoniae transferrin receptor genes

Characterization of a highly structured domain in Tbp2 from Neisseria meningitidis involved in binding to human transferrin

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