A general strategy for the assignment of aliphatic side-chain resonances of uniformly 13C,15N-labeled large proteins.
A general strategy is proposed to assign aliphatic side-chain resonances of large 13C,15N-labeled proteins without deuteration, using 4D 13C,15N-edited NOESY and MQ-(H)CCH-TOCSY experiments on the basis of prior assignments of backbone and 13Cbeta resonances. The strategy has been tested on a 214 residue protein (DdCAD-1) and applied to a chain-selectively 13C,15N-labeled hemoglobin (65 kDa). About 96 and 80% aliphatic side-chain spins in DdCAD-1 and hemoglobin have been assigned, respectively. The strategy proposed here will be very useful for the structure determination and dynamics characterization of large proteins by NMR.
Xu
Yingqi
Y
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543.
Lin
Zhi
Z
Ho
Chien
C
Yang
Daiwen
D
D002247:Carbon Isotopes / Q000737:chemistry*; D006839:Hydrocarbons, Acyclic / Q000737:chemistry*; D008970:Molecular Weight; D009587:Nitrogen Isotopes / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
D002247:Carbon Isotopes; D006839:Hydrocarbons, Acyclic; D009587:Nitrogen Isotopes; D011506:Proteins