Sequence analysis Analyses of DNA and protein sequences and design of oligonucleotides were facilitated by the Lasergene software package of DNA star Inc. (Madison, Wis.). Homology searches were done by Blast analysis http://blast.ncbi.nlm.nih.gov. In silico secondary structure analyses of the OppA variants were performed by the SOPMA Secondary Prediction Method (Pôle BioInformatique Lyonnaise network proteon sequence analysis; http://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html)
Statistical analysis All experiments were performed in triplicate, with similar CP673451 mouse results obtained by at least three independent tests. Km and Vmax were calculated with a computerized nonlinear regression analysis (Graph Pad Prism, version 5.01; Graph Pad Software Inc. Sang Diego, Calif.). Funding This work was supported by a grant from the research commission of the medical faculty of the Heinrich-Heine University Duesseldorf, Germany. Acknowledgements Peptide 17 supplier We thank Dana Belick for excellent technical assistance, especially for tireless purifications of the recombinant
OppA mutants. We are indebted to Heiner Schaal for his helpful discussion of the manuscript, as well as Colin MacKenzie and Elisabeth Kravets for critically reading the manuscript. References 1. Kline KA, Falker S, Dahlberg S, Normark S, Henriques-Normark B: Bacterial Adhesins in Host-Microbe Interactions. Cell Host & Microbe 2009, 5:580–592.CrossRef 2. Kawahito Y, Ichinose S, Sano H, Tsubouchi Y, Kohno M, AZD6244 Yoshikawa T, Tokunaga D, Hojo T, Harasawa R, Nakano ID-8 T, Matsuda K: Mycoplasma fermentans glycolipid-antigen as a pathogen of rheumatoid arthritis. Biochem Biophys Res Commun 2008, 369:561–566.PubMedCrossRef 3. Rottem S: Choline-containing lipids in mycoplasmas.
Microbes Infect 2002, 4:963–968.PubMedCrossRef 4. Yavlovich A, Katzenell A, Tarshis M, Higazi AAR, Rottem S: Mycoplasma fermentans binds to and invades HeLa cells: Involvement of plasminogen and urokinase. Infect Immun 2004, 72:5004–5011.PubMedCrossRef 5. Berg M, Melcher U, Fletcher J: Characterization of Spiroplasma citri adhesion related protein SARP1, which contains a domain of a novel family designated sarpin 1. Gene 2001, 275:57–64.PubMedCrossRef 6. Henrich B, Feldmann RC, Hadding U: Cytoadhesins of Mycoplasma hominis. Infect Immun 1993, 61:2945–2951.PubMed 7. Djordjevic SP, Cordwell SJ, Djordjevic MA, Wilton J, Minion FC: Proteolytic processing of the Mycoplasma hyopneumoniae cilium adhesin. Infect Immun 2004, 72:2791–2802.PubMedCrossRef 8. Leigh SA, Wise KS: Identification and functional mapping of the mycoplasma fermentans P29 adhesin. Infect Immun 2002, 70:4925–4935.PubMedCrossRef 9.