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  • 1
    Electronic Resource
    Electronic Resource
    Measurement of amide proton exchange rates and NOEs with water in 13C/15N-enriched calcineurin B (1993)
    Springer
    Journal of biomolecular NMR 3 (1993), S. 627-638 
    Details
    ISSN: 1573-5001
    Keywords: Hydrogen exchange ; NOESY ; ROESY ; Water ; Protein ; 2D NMR ; Calcineurin B
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary A rapid and sensitive 2D approach is presented for measuring amide proton exchange rates and the NOE interaction between amide protons and water. The approach is applicable to uniformly 13C/15N-enriched proteins and can measure magnetization exchange rates in the 0.02 to 〉20s−1 range. The experiments rely on selective excitation of the water resonance, coupled with purging of underlying Hα resonances, followed by NOESY-or ROESY-type transfer to amide protons, which are dispersed by the amide 15N frequencies in an HSQC-type experiment. Two separate but interleaved experiments, with and without selective inversion of the H2O resonance, yield quantitative results. The method is demonstrated for a sample of the calcium-binding protein calcineurin B. Results indicate rapid amide exchange for the five calcineurin B residues that are analogous to the five rapidly exchanging residues in the ‘central helix’ of the homologous protein calmodulin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00198368
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  • 2
    Electronic Resource
    Electronic Resource
    Spin-locked multiple quantum coherence for signal enhancement in heteronuclear multidimensional NMR experiments (1995)
    Springer
    Journal of biomolecular NMR 6 (1995), S. 335-339 
    Details
    ISSN: 1573-5001
    Keywords: Multidimensional NMR ; Line narrowing ; Relaxation ; Multiple quantum coherence ; ROESY ; Protein ; Calmodulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary For methine sites the relaxation rate of 13C-1H two-spin coherence is generally slower than the relaxation rate of the individual 13C and 1H single spin coherences. The slower decay of two-spin coherence can be used to increase the sensitivity and resolution in heteronuclear experiments, particularly those that require correlation of Hα and Cα chemical shifts. To avoid dephasing of the two-spin coherence caused by 1H-1H J-couplings, the 1H spin is locked by the application of a weak rf field, resulting in a spin-locked multiple quantum coherence. For a sample of calcium-free calmodulin, use of the multiple quantum approach yields significant signal enhancement over the conventional constant-time 2D HSQC experiment. The approach is applicable to many multidimensional NMR experiments, as demonstrated for a 3D 13C-separated ROESY CT-HMQC spectrum.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00197815
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