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P80, the HinT interacting membrane protein, is a secreted antigen of Mycoplasma hominis

BioMed Central
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Abstract ral ss BioMed CentBMC Microbiology Open AcceResearch article P80, the HinT interacting membrane protein, is a secreted antigen of Mycoplasma hominis Miriam Hopfe, Ricarda Hoffmann and Birgit Henrich* Address: Institute of Medical Microbiology and Center for Biological and Medical Research, Heinrich-Heine-University, Moorenstrasse 5, 40225 Duesseldorf, Germany Email: Miriam Hopfe - [email protected]; Ricarda Hoffmann - [email protected]; Birgit Henrich* - [email protected] * Corresponding author Abstract Background: Mycoplasmas are cell wall-less bacteria which encode a minimal set of proteins. In Mycoplasma hominis, the genes encoding the surface-localized membrane complex P60/P80 are in an operon with a gene encoding a cytoplasmic, nucleotide-binding protein with a characteristic Histidine triad motif (HinT). HinT is found in both procaryotes and eukaryotes and known to hydrolyze adenosine nucleotides in eukaryotes. Immuno-precipitation and BIACore analysis revealed an interaction between HinT and the P80 domain of the membrane complex. As the membrane anchored P80 carries an N-terminal uncleaved signal peptide we have proposed that the N-terminus extends into the cytoplasm and interacts with the cytosolic HinT. Results: Further characterization of P80 suggested that the 4.7 kDa signal peptide is protected from cleavage only in the membrane bound form. We found several proteins were released into the supernatant of a logarithmic phase mycoplasma culture, including P80, which was reduced in size by 10 kDa. Western blot analysis of recombinant P80 mutants expressed in E. coli and differing in the N-terminal region revealed that mutation of the +1 position of the mature protein (Asn to Pro) which is important for signal peptidase I recognition resulted in reduced P80 secretion. All other P80 variants were released into the supernatant, in general as a 74 kDa protein encompassing the helical part of P80. Incubation of M. homi

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