caa a22 4500 445325801 CHVBK 20180317142720.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s10858-011-9511-7 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9511-7 Sugase Kenji Bioorganic Research Institute, Suntory Foundation for Life Sciences, 1-1-1 Wakayamadai, Shimamoto-cho, 618-8503, Mishima-gun, Osaka, Japan aut Elucidating slow binding kinetics of a protein without observable bound resonances by longitudinal relaxation NMR spectroscopy [Elektronische Daten] [Kenji Sugase] We developed a new method to elucidate the binding kinetics kon and koff, and the dissociation constant KD (=koff/kon), of protein-protein interactions without observable bound resonances of the protein of interest due to high molecular weight in a complex with a large target protein. In our method, kon and koff rates are calculated from the analysis of longitudinal relaxation rates of free resonances measured for multiple samples containing different concentration ratios of 15N-labeled protein and substoichiometric amounts of the target protein. The method is applicable to interactions that cannot be analyzed by relaxation dispersion spectroscopy due to slow interactions on millisecond to second timescale and/or minimal conformational (chemical shift) change upon binding. We applied the method to binding of the B1 domain of protein G (GB1) to immunoglobulin G, and derived the binding kinetics despite the absence of observable bound GB1 resonances. Springer Science+Business Media B.V., 2011 Binding kinetics nationallicence Dissociation constant nationallicence Longitudinal relaxation experiment nationallicence Slow exchange nationallicence Unobservable bound resonance nationallicence Journal of Biomolecular NMR Springer Netherlands 50/3(2011-07-01), 219-227 0925-2738 50:3<219 2011 50 10858 https://doi.org/10.1007/s10858-011-9511-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9511-7 text/html Onlinezugriff via DOI NATIONALLICENCE 100 1- Sugase Kenji Bioorganic Research Institute, Suntory Foundation for Life Sciences, 1-1-1 Wakayamadai, Shimamoto-cho, 618-8503, Mishima-gun, Osaka, Japan aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 50/3(2011-07-01), 219-227 0925-2738 50:3<219 2011 50 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer