Dear Editor,The functional diversity of proteins is related to the cooperation of multiple domains.Independent globular domains are typically joined by a fl exible length of polypeptide chain,which makes the structura...Dear Editor,The functional diversity of proteins is related to the cooperation of multiple domains.Independent globular domains are typically joined by a fl exible length of polypeptide chain,which makes the structural analysis of multi-domain proteins diffi cult.Here,we describe the combined use of solution NMR(nuclear magnetic resonance)and EPR(elec-tron paramagnetic resonance)for the structural analysis of a protein with two separate domains.The structure of each domain was determined independently using conventional NMR restraints,and the relative orientation of the two separate domains was confi ned using long-distance restraints obtained by NMR-PRE(paramagnetic relaxation enhancement)and EPR-DEER(double electron-electron resonance,also called PELDOR:pulsed electron double reso-nance.展开更多
Dear Editor, Accurate measurement of distances within and between biological macromolecules is important for structure-function studies, for investigating conformational changes and dynamics, and for improving restrai...Dear Editor, Accurate measurement of distances within and between biological macromolecules is important for structure-function studies, for investigating conformational changes and dynamics, and for improving restraints for molecular dynamics and modeling programs （Hellmich and Glaubitz, 2009）. Several methods have been developed to measure the distances between atoms, residues, and domains in single proteins and larger complexes. Fluorescence reso- nance energy transfer （FRET） is currently the most fre- quently used method to derive distances in cells or protein complexes, but the bulky size of the fluorescence probes and the strict requirement for frequency profile overlap between the acceptor and donor chromophores are limita- tions in many cases （Prevo and Peterman, 2014）.展开更多
基金This research was supported by the National Basic Research Program(973 Program)(Nos.2011CB911104 and 2013CB910200)the Chinese Natural Science Foundation of China(Grant No.31100563)to Y.Xiong and(Grant No.31170817)to C.Tian.
文摘Dear Editor,The functional diversity of proteins is related to the cooperation of multiple domains.Independent globular domains are typically joined by a fl exible length of polypeptide chain,which makes the structural analysis of multi-domain proteins diffi cult.Here,we describe the combined use of solution NMR(nuclear magnetic resonance)and EPR(elec-tron paramagnetic resonance)for the structural analysis of a protein with two separate domains.The structure of each domain was determined independently using conventional NMR restraints,and the relative orientation of the two separate domains was confi ned using long-distance restraints obtained by NMR-PRE(paramagnetic relaxation enhancement)and EPR-DEER(double electron-electron resonance,also called PELDOR:pulsed electron double reso-nance.
文摘Dear Editor, Accurate measurement of distances within and between biological macromolecules is important for structure-function studies, for investigating conformational changes and dynamics, and for improving restraints for molecular dynamics and modeling programs （Hellmich and Glaubitz, 2009）. Several methods have been developed to measure the distances between atoms, residues, and domains in single proteins and larger complexes. Fluorescence reso- nance energy transfer （FRET） is currently the most fre- quently used method to derive distances in cells or protein complexes, but the bulky size of the fluorescence probes and the strict requirement for frequency profile overlap between the acceptor and donor chromophores are limita- tions in many cases （Prevo and Peterman, 2014）.
