caa a22 4500 477135781 CHVBK 20180405111717.0 cr unu---uuuuu 170330e19970701xx s 000 0 eng 10.1023/A:1018339526108 doi (NATIONALLICENCE)springer-10.1023/A:1018339526108 Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR [Elektronische Daten] [Andrew Dingley, Joel Mackay, Graeme Shaw, Brett Hambly, Glenn King] We have previously shown that 1H pulsed-field-gradient(PFG) NMR spectroscopy provides a facile method for monitoring proteinself-association and can be used, albeit with some caveats, to measure theapparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol.NMR, 6, 321-328]. In this paper we show that, for15N-labelled proteins, selection of1H-15N multiple-quantum (MQ) coherences in PFGdiffusion experiments provides several advantages over monitoring1H single-quantum (SQ) magnetization. First, the use of agradient-selected MQ filter provides a convenient means of suppressingresonances from both the solvent and unlabelled solutes. Second,1H-15N zero-quantum coherence dephases morerapidly than 1H SQ coherence under the influence of a PFG.This allows the diffusion coefficients of larger proteins to be measuredmore readily. Alternatively, the gradient length and/or the diffusion delaymay be decreased, thereby reducing signal losses from relaxation. In orderto extend the size of macromolecules to which these experiments can beapplied, we have developed a new MQ PFG diffusion experiment in which themagnetization is stored as longitudinal two-spin order for most of thediffusion period, thus minimizing sensitivity losses due to transverserelaxation and J-coupling evolution. Kluwer Academic Publishers, 1997 Pulsed-field-gradient NMR nationallicence Translational diffusion coefficient nationallicence Self-association nationallicence Macromolecules nationallicence Solvent suppression nationallicence Dingley Andrew Department of Biochemistry, University of Sydney, 2006, Sydney, NSW, Australia aut Mackay Joel Department of Biochemistry, University of Sydney, 2006, Sydney, NSW, Australia aut Shaw Graeme Department of Biochemistry, University of Oxford, OX1 3QU, Oxford, U.K aut Hambly Brett Department of Pathology, University of Sydney, 2006, Sydney, NSW, Australia aut King Glenn Department of Biochemistry, University of Sydney, 2006, Sydney, NSW, Australia aut Journal of Biomolecular NMR Kluwer Academic Publishers 10/1(1997-07-01), 1-8 0925-2738 10:1<1 1997 10 10858 https://doi.org/10.1023/A:1018339526108 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018339526108 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dingley Andrew Department of Biochemistry, University of Sydney, 2006, Sydney, NSW, Australia aut NATIONALLICENCE 700 1- Mackay Joel Department of Biochemistry, University of Sydney, 2006, Sydney, NSW, Australia aut NATIONALLICENCE 700 1- Shaw Graeme Department of Biochemistry, University of Oxford, OX1 3QU, Oxford, U.K aut NATIONALLICENCE 700 1- Hambly Brett Department of Pathology, University of Sydney, 2006, Sydney, NSW, Australia aut NATIONALLICENCE 700 1- King Glenn Department of Biochemistry, University of Sydney, 2006, Sydney, NSW, Australia aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 10/1(1997-07-01), 1-8 0925-2738 10:1<1 1997 10 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer