Pulse schemes for the measurement of 3JC'C gamma and 3JNC gamma scalar couplings in 15N,13C uniformly labeled proteins.
Pulse sequences are presented for the measurement of 3JC'C gamma and 3JNC gamma scalar couplings for all C gamma containing residues in 15N,13C uniformly labeled proteins. The methods described are based on quantitative J correlation spectroscopy pioneered by Bax and co-workers [Bax et. al. (1994) Methods Enzymol., 239, 79-105]. The combination of 3JC'C gamma and 3JNC gamma scalar coupling constants allows the assignment of discrete rotameric states about the chi 1 torsion angle in cases where such states exist or, alternatively, facilitates the establishment of noncanonical chi 1 conformations or the presence of rotameric averaging. The methods are applied to a 1.5 mM sample of staphylococcal nuclease.
Konrat
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Protein Engineering Network of Centres of Excellence, University of Toronto, ON, Canada.
Muhandiram
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DR
Farrow
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Kay
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LE
D000596:Amino Acids / Q000737:chemistry; D002247:Carbon Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008836:Micrococcal Nuclease / Q000737:chemistry*; D008956:Models, Chemical; D009587:Nitrogen Isotopes; D011487:Protein Conformation
D000596:Amino Acids; D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D008836:Micrococcal Nuclease