دورية أكاديمية
NMR exchange broadening arising from specific low affinity protein self-association: analysis of nitrogen-15 nuclear relaxation for rat CD2 domain 1.
العنوان: | NMR exchange broadening arising from specific low affinity protein self-association: analysis of nitrogen-15 nuclear relaxation for rat CD2 domain 1. |
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المؤلفون: | Pfuhl M; Department of Biochemistry and Molecular Biology, University College London, U.K., Chen HA, Kristensen SM, Driscoll PC |
المصدر: | Journal of biomolecular NMR [J Biomol NMR] 1999 Aug; Vol. 14 (4), pp. 307-20. |
نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: Springer Country of Publication: Netherlands NLM ID: 9110829 Publication Model: Print Cited Medium: Print ISSN: 0925-2738 (Print) Linking ISSN: 09252738 NLM ISO Abbreviation: J Biomol NMR Subsets: MEDLINE |
أسماء مطبوعة: | Publication: <2005->: Leiden, The Netherlands : Springer Original Publication: Leiden, The Netherlands : ESCOM Science Publishers, c1991- |
مواضيع طبية MeSH: | CD2 Antigens/*chemistry, Animals ; Dimerization ; Magnetic Resonance Spectroscopy ; Nitrogen ; Rats |
مستخلص: | Nuclear spin relaxation monitored by heteronuclear NMR provides a useful method to probe the overall and internal molecular motion for biological macromolecules over a variety of time scales. Nitrogen-15 NMR relaxation parameters have been recorded for the N-terminal domain of the rat T-cell antigen CD2 (CD2d1) in a dilution series from 1.20 mM to 40 microM (pH 6.0, 25 degrees C). The data have been analysed within the framework of the model-free formalism of Lipari and Szabo to understand the molecular origin of severely enhanced transverse relaxation rates found for certain residues. These data revealed a strong dependence of the derived molecular correlation time tau c upon the CD2d1 protein concentration. Moreover, a number of amide NH resonances exhibited exchange broadening and chemical shifts both strongly dependent on protein concentration. These amide groups cluster on the major beta-sheet surface of CD2d1 that coincides with a major lattice contact in the X-ray structure of the intact ectodomain of rat CD2. The complete set of relaxation data fit well to an equilibrium monomer-dimer exchange model, yielding estimates of exchange rate constants (kON = 5000 M-1 s-1; kOFF = 7 s-1) and a dissociation constant (KD approximately 3-6 mM) that is consistent with the difficulty in detecting the weak interactions for this molecule by alternative biophysical methods. The self-association of CD2d1 is essentially invariant to changes in buffer composition and ionic strength and the associated relaxation phenomena cannot be explained as a result of neglecting anisotropic rotational diffusion in the analysis. These observations highlight the necessity to consider low affinity protein self-association interactions as a source of residue specific exchange phenomena in NMR spectra of macromolecular biomolecules, before the assignment of more elaborate intramolecular conformational mechanisms. |
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المشرفين على المادة: | 0 (CD2 Antigens) N762921K75 (Nitrogen) |
تواريخ الأحداث: | Date Created: 19991020 Date Completed: 19991102 Latest Revision: 20190921 |
رمز التحديث: | 20231215 |
DOI: | 10.1023/a:1008319917267 |
PMID: | 10526406 |
قاعدة البيانات: | MEDLINE |
تدمد: | 0925-2738 |
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DOI: | 10.1023/a:1008319917267 |