caa a22 4500 445325615 CHVBK 20180317142719.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s10858-011-9498-0 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9498-0 Measuring 1HN temperature coefficients in invisible protein states by relaxation dispersion NMR spectroscopy [Elektronische Daten] [Guillaume Bouvignies, Pramodh Vallurupalli, Matthew Cordes, D. Hansen, Lewis Kay] A method based on the Carr-Purcell-Meiboom-Gill relaxation dispersion experiment is presented for measuring the temperature coefficients of amide proton chemical shifts of low populated ‘invisible' protein states that exchange with a ‘visible' ground state on the millisecond time-scale. The utility of the approach is demonstrated with an application to an I58D mutant of the Pfl6 Cro protein that undergoes exchange between the native, folded state and a cold denatured, unfolded conformational ensemble that is populated at a level of 6% at 2.5°C. A wide distribution of amide temperature coefficients is measured for the unfolded state. The distribution is centered about -5.6ppb/K, consistent with an absence of intra-molecular hydrogen bonds, on average. However, the large range of values (standard deviation of 2.1ppb/K) strongly supports the notion that the unfolded state of the protein is not a true random coil polypeptide chain. Springer Science+Business Media B.V., 2011 CPMG relaxation dispersion nationallicence Temperature coefficients nationallicence Amide protons nationallicence Cold denaturation nationallicence Bouvignies Guillaume Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut Vallurupalli Pramodh Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut Cordes Matthew Department of Chemistry and Biochemistry, The University of Arizona, 85721-0088, Tucson, AZ, USA aut Hansen D. Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut Kay Lewis Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut Journal of Biomolecular NMR Springer Netherlands 50/1(2011-05-01), 13-18 0925-2738 50:1<13 2011 50 10858 https://doi.org/10.1007/s10858-011-9498-0 text/html Onlinezugriff via DOI 1 brief-communication jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9498-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bouvignies Guillaume Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Vallurupalli Pramodh Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Cordes Matthew Department of Chemistry and Biochemistry, The University of Arizona, 85721-0088, Tucson, AZ, USA aut NATIONALLICENCE 700 1- Hansen D. Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Kay Lewis Departments of Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, M5S 1A8, Toronto, ON, Canada aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 50/1(2011-05-01), 13-18 0925-2738 50:1<13 2011 50 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer