The Adherence-Associated Lipoprotein P100, Encoded by an opp Operon Structure, Functions as the Oligopeptide-Binding Domain OppA of a Putative Oligopeptide Transport System in Mycoplasma hominis
| Autor: | Ulrich Hadding, Miriam Hopfe, Annette Kitzerow, Birgit Henrich |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
genetic structures
Operon Lipoproteins Molecular Sequence Data Cell Surfaces Mycoplasma hominis Biology Oligopeptide transport Microbiology Bacterial Adhesion Bacterial Proteins Amino Acid Sequence RNA Messenger Molecular Biology Peptide sequence Gene Genetics Base Sequence Membrane transport protein Chromosome Mapping Membrane Transport Proteins Biological Transport Blotting Northern biology.organism_classification Molecular biology RNA Bacterial Spectrometry Fluorescence Peptide transport biology.protein Carrier Proteins Oligonucleotide Probes Oligopeptides Oligopeptide binding Plasmids |
| Zdroj: | Journal of Bacteriology. 181:4873-4878 |
| ISSN: | 1098-5530 0021-9193 |
| Popis: | Mycoplasma hominis, a cell-wall-less prokaryote, was shown to be cytoadherent by the participation of a 100-kDa membrane protein (P100). To identify the gene encoding P100, peptides of P100 were partially sequenced to enable the synthesis of P100-specific oligonucleotides suitable as probes for the detection of the P100 gene. With this strategy, we identified a genomic region of about 10.4 kb inM. hominisFBG carrying the P100 gene. Analysis of the complete deduced protein sequence suggests that P100 is expressed as a pre-lipoprotein with a structure in the N-terminal region common to peptide-binding proteins and an ATP- or GTP-binding P-loop structure in the C-terminal region. Downstream of the P100 gene, an additional four open reading frames putatively encoding the four core domains of an active transport system, OppBCDF, were localized. The organization of the P100 gene andoppBCDFin a transcriptionally active operon structure was demonstrated in Northern blot and reverse transcription-PCR analyses, as all gene-specific probes detected a common RNA of 9.5 kb. Primer extension analysis revealed that the transcriptional initiation site was localized 323 nucleotides upstream of the methionine-encoding ATG of the P100 gene. The peptide-binding character of the P100 protein was confirmed by fluorescence spectroscopy and strongly suggests that the cytoadherence-mediating lipoprotein P100 represents OppA, the substrate-binding domain of a peptide transport system inM. hominis. |
| Databáze: | OpenAIRE |
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