Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2. Dingley, A., Mackay, J., Chapman, B., Morris, M., Kuchel, P., Hambly, B., & King, G. Journal of Biomolecular NMR, 6(3):321-328, 11, 1995.
Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2 [link]Website  abstract   bibtex   
At the millimolar concentrations required for structural studies, NMR spectra of the calcium-binding protein myosin light chain 2 (MLC2) showed resonance line widths indicative of extensive self-association. Pulsed-field-gradient (PFG) NMR spectroscopy was used to examine whether MLC2 aggregation could be prevented by the zwitterionic bile salt derivative 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). PFG NMR measurements indicated that CHAPS was capable of preventing MLC2 self-association, but only at concentrations well above the critical micelle concentration of ∼7.5 mM. CHAPS was most effective at a concentration of 22.5 mM, where the apparent molecular mass of MLC2 correponded to a protein monomer plus seven molecules of bound detergent. The resolution and sensitivity of 2D 15N-1H HSQC spectra of MLC2 were markedly improved by the addition of 25 mM CHAPS, consistent with a reduction in aggregation following addition of the detergent. The average amide nitrogen T2 value for MLC2 increased from ∼30 ms in the absence of CHAPS to ∼56 ms in the presence of 25 mM CHAPS. The results of this study lead us to propose that PFG NMR spectroscopy can be used as a facile alternative to conventional techniques such as analytical ultracentrifugation for examining the self-association of biological macromolecules. © 1995 ESCOM Science Publishers B.V.
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 title = {Measuring protein self-association using pulsed-field-gradient NMR spectroscopy: Application to myosin light chain 2},
 type = {article},
 year = {1995},
 identifiers = {[object Object]},
 keywords = {CHAPS,Myosin light chain,Pulsed-field-gradient NMR,Self-association,Translational diffusion coefficient},
 pages = {321-328},
 volume = {6},
 websites = {http://link.springer.com/10.1007/BF00197813},
 month = {11},
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 abstract = {At the millimolar concentrations required for structural studies, NMR spectra of the calcium-binding protein myosin light chain 2 (MLC2) showed resonance line widths indicative of extensive self-association. Pulsed-field-gradient (PFG) NMR spectroscopy was used to examine whether MLC2 aggregation could be prevented by the zwitterionic bile salt derivative 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). PFG NMR measurements indicated that CHAPS was capable of preventing MLC2 self-association, but only at concentrations well above the critical micelle concentration of ∼7.5 mM. CHAPS was most effective at a concentration of 22.5 mM, where the apparent molecular mass of MLC2 correponded to a protein monomer plus seven molecules of bound detergent. The resolution and sensitivity of 2D 15N-1H HSQC spectra of MLC2 were markedly improved by the addition of 25 mM CHAPS, consistent with a reduction in aggregation following addition of the detergent. The average amide nitrogen T2 value for MLC2 increased from ∼30 ms in the absence of CHAPS to ∼56 ms in the presence of 25 mM CHAPS. The results of this study lead us to propose that PFG NMR spectroscopy can be used as a facile alternative to conventional techniques such as analytical ultracentrifugation for examining the self-association of biological macromolecules. © 1995 ESCOM Science Publishers B.V.},
 bibtype = {article},
 author = {Dingley, AndrewJ. and Mackay, JoelP. and Chapman, BogdanE. and Morris, MichaelB. and Kuchel, PhilipW. and Hambly, BrettD. and King, GlennF.},
 journal = {Journal of Biomolecular NMR},
 number = {3}
}

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