caa a22 4500 605505977 CHVBK 20210128100624.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s00253-015-6456-x doi (NATIONALLICENCE)springer-10.1007/s00253-015-6456-x In vitro effect of temperature on the conformational structure and collagen binding of SdrF, a Staphylococcus epidermidis adhesin [Elektronische Daten] [Antonella Di Poto, Massimiliano Papi, Sheetal Trivedi, Alessandro Maiorana, Paola Gavazzo, Massimo Vassalli, Franklin Lowy, Marco De Spirito, Lucio Montanaro, Marcello Imbriani, Carla Arciola, Livia Visai] Staphylococcus epidermidis is the leading etiologic agent of device-related infections. S. epidermidis is able to bind, by means of the adhesins of its cell wall, the host matrix proteins filming the artificial surfaces. Thence, bacteria cling to biomaterials and infection develops. The effect of temperature on integrity, structure, and biological activity of the collagen-binding adhesin (SdrF) of S. epidermidis has been here investigated. By cloning in E. coli XL1-Blue, a recombinant of the SdrF binding domain B (rSdrFB), carrying an N-terminal polyhistidine, was obtained. Purification was by HiTrapTM Chelating HP columns. Assessment of purity, molecular weight, and integrity was by SDS-PAGE. The rSdrFB-collagen binding was investigated by ELISA. A full three-dimensional reconstruction of rSdrFB was achieved by small-angle X-ray scattering (SAXS). At 25°C, rSdrFB bound to type I collagen in a dose-dependent, saturable manner, with a Kd of 2.48 × 10−7M. When temperature increased from 25 to 37°C, a strong conformational change occurred, together with the abolition of the rSdrFB-collagen binding. The rSdrFB integrity was not affected by temperature variation. SdrFB-collagen binding is switched on/off depending on the temperature. Implications with the infection pathogenesis are enlightened. Springer-Verlag Berlin Heidelberg, 2015 Staphylococcus epidermidis nationallicence SdrF adhesin nationallicence Collagen nationallicence Small-angle X-ray scattering (SAXS) nationallicence Normalized spatial discrepancies (NSD) nationallicence Hypothermia nationallicence Opportunistic infections nationallicence Di Poto Antonella Department of Molecular Medicine, Center for Tissue Engineering (CIT), INSTM UdR of Pavia, University of Pavia, Viale Taramelli 3/b, 27100, Pavia, Italy aut Papi Massimiliano Physics Institute, Sacred Heart Catholic University, Largo F. Vito, 1, 00168, Rome, Italy aut Trivedi Sheetal Division of Infectious Diseases, Department of Medicine, College of Physicians and Surgeons, Columbia University, 630 West 168th Street Box 82, 10032, New York, NY, USA aut Maiorana Alessandro Physics Institute, Sacred Heart Catholic University, Largo F. Vito, 1, 00168, Rome, Italy aut Gavazzo Paola Institute of Biophysics of the National Research Council of Italy, Via De Marini 6, 16149, Genoa, Italy aut Vassalli Massimo Institute of Biophysics of the National Research Council of Italy, Via De Marini 6, 16149, Genoa, Italy aut Lowy Franklin Division of Infectious Diseases, Department of Medicine, College of Physicians and Surgeons, Columbia University, 630 West 168th Street Box 82, 10032, New York, NY, USA aut De Spirito Marco Physics Institute, Sacred Heart Catholic University, Largo F. Vito, 1, 00168, Rome, Italy aut Montanaro Lucio Research Unit on Implant Infections, Rizzoli Orthopedic Institute, via di Barbiano 1/10, 40136, Bologna, Italy aut Imbriani Marcello Department of Public Health, Experimental Medicine and Forensics, University of Pavia, Via Forlanini 2, 27100, Pavia, Italy aut Arciola Carla Research Unit on Implant Infections, Rizzoli Orthopedic Institute, via di Barbiano 1/10, 40136, Bologna, Italy aut Visai Livia Department of Molecular Medicine, Center for Tissue Engineering (CIT), INSTM UdR of Pavia, University of Pavia, Viale Taramelli 3/b, 27100, Pavia, Italy aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5593-5603 0175-7598 99:13<5593 2015 99 253 https://doi.org/10.1007/s00253-015-6456-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6456-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Di Poto Antonella Department of Molecular Medicine, Center for Tissue Engineering (CIT), INSTM UdR of Pavia, University of Pavia, Viale Taramelli 3/b, 27100, Pavia, Italy aut NATIONALLICENCE 700 1- Papi Massimiliano Physics Institute, Sacred Heart Catholic University, Largo F. Vito, 1, 00168, Rome, Italy aut NATIONALLICENCE 700 1- Trivedi Sheetal Division of Infectious Diseases, Department of Medicine, College of Physicians and Surgeons, Columbia University, 630 West 168th Street Box 82, 10032, New York, NY, USA aut NATIONALLICENCE 700 1- Maiorana Alessandro Physics Institute, Sacred Heart Catholic University, Largo F. Vito, 1, 00168, Rome, Italy aut NATIONALLICENCE 700 1- Gavazzo Paola Institute of Biophysics of the National Research Council of Italy, Via De Marini 6, 16149, Genoa, Italy aut NATIONALLICENCE 700 1- Vassalli Massimo Institute of Biophysics of the National Research Council of Italy, Via De Marini 6, 16149, Genoa, Italy aut NATIONALLICENCE 700 1- Lowy Franklin Division of Infectious Diseases, Department of Medicine, College of Physicians and Surgeons, Columbia University, 630 West 168th Street Box 82, 10032, New York, NY, USA aut NATIONALLICENCE 700 1- De Spirito Marco Physics Institute, Sacred Heart Catholic University, Largo F. Vito, 1, 00168, Rome, Italy aut NATIONALLICENCE 700 1- Montanaro Lucio Research Unit on Implant Infections, Rizzoli Orthopedic Institute, via di Barbiano 1/10, 40136, Bologna, Italy aut NATIONALLICENCE 700 1- Imbriani Marcello Department of Public Health, Experimental Medicine and Forensics, University of Pavia, Via Forlanini 2, 27100, Pavia, Italy aut NATIONALLICENCE 700 1- Arciola Carla Research Unit on Implant Infections, Rizzoli Orthopedic Institute, via di Barbiano 1/10, 40136, Bologna, Italy aut NATIONALLICENCE 700 1- Visai Livia Department of Molecular Medicine, Center for Tissue Engineering (CIT), INSTM UdR of Pavia, University of Pavia, Viale Taramelli 3/b, 27100, Pavia, Italy aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5593-5603 0175-7598 99:13<5593 2015 99 253