Sequence-specific assignments of methyl groups in high-molecular weight proteins.
A new 3D multiple-quantum (H)CCmHm-TOCSY experiment is proposed to assign methyl resonances in high-molecular weight proteins, on the basis of spectral patterns and prior backbone assignments. The favorable relaxation properties of the multiple-quantum coherences and the slow decays of in-phase methyl 13C magnetizations optimize performance of the proposed experiment for application to large proteins. The experiment has been demonstrated on an acyl carrier protein synthase (trimer, 42 kDa, overall correlation time of 26 ns) at 25 degrees C, and 63 out of 67 nonmethionine methyl groups have been assigned.
Yang
Daiwen
D
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543. dbsydw@nus.edu.sg
Zheng
Yu
Y
Liu
Dingjiang
D
Wyss
Daniel F
DF
D002247:Carbon Isotopes; D002352:Carrier Proteins / Q000737:chemistry*; D008970:Molecular Weight; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D011522:Protons; D017855:Transferases (Other Substituted Phosphate Groups) / Q000737:chemistry
D002247:Carbon Isotopes; D002352:Carrier Proteins; D009587:Nitrogen Isotopes; D011522:Protons; D017855:Transferases (Other Substituted Phosphate Groups); C021364:holo-(acyl-carrier-protein) synthase