Protein-protein interactions （PPI） are essential for a variety of cellular functions. Many PPI analyses were conducted in vitro, using purified proteins. In this report, the unnatural amino acid tfmF was site-specif...Protein-protein interactions （PPI） are essential for a variety of cellular functions. Many PPI analyses were conducted in vitro, using purified proteins. In this report, the unnatural amino acid tfmF was site-specifically incorporated into several different sites of two Phox-Beml （PB1） domains from two mitogen activated protein kinases （MEKK3 and/ or MEK5） in the E. coil cells.展开更多
Current resolved structures of GPCRs and G protein complexes provided important insights into G protein activation. However, the binding or dissociation of GPCRs with G protein is instantaneous and highly dynamic in t...Current resolved structures of GPCRs and G protein complexes provided important insights into G protein activation. However, the binding or dissociation of GPCRs with G protein is instantaneous and highly dynamic in the intracellular environment. The conformational dynamic of G protein still needs to be addressed. In this study, we applied ^(19)F solution NMR spectroscopy to monitor the conformational changes of G protein upon interact with detergent mimicking membrane and receptor. Our results show that there are two states equilibria in the G_(α)in apo states. The interaction of G_(α)with detergents will accelerate this conformational transformation and induce a state that tends to bind to GPCRs. Finally, the G_(α)proteins presented a fully activation state when they coupled to GPCRs.展开更多
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
ERRATUM TO： PROTEIN CELL 2017, 8（2）：149-154 DOI 10.1007/S13238-016-0336-8In the original publication of this article the correspondence authors＇ e-mail has been missed out. The missing e-mail addresses are prov...ERRATUM TO： PROTEIN CELL 2017, 8（2）：149-154 DOI 10.1007/S13238-016-0336-8In the original publication of this article the correspondence authors＇ e-mail has been missed out. The missing e-mail addresses are provided in this erratum.OPEN ACCESS This article is distributed under the terms of the Creative Commons Attribution 4.0 international License （http：//creativecommons.org/ licenses/by/4.0/）, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author（s） and the source, provide a link to the Creative Commons license, and indicate if changes were made.展开更多
文摘Protein-protein interactions （PPI） are essential for a variety of cellular functions. Many PPI analyses were conducted in vitro, using purified proteins. In this report, the unnatural amino acid tfmF was site-specifically incorporated into several different sites of two Phox-Beml （PB1） domains from two mitogen activated protein kinases （MEKK3 and/ or MEK5） in the E. coil cells.
基金supported by the National Key Research and Development Project of China (Nos.2019YFA0904100 and 2017YFA0505400)the National Natural Science Foundation of China (Nos.22077117 and 31971152)the USTC Research Funds of the Double First-Class Initiative。
文摘Current resolved structures of GPCRs and G protein complexes provided important insights into G protein activation. However, the binding or dissociation of GPCRs with G protein is instantaneous and highly dynamic in the intracellular environment. The conformational dynamic of G protein still needs to be addressed. In this study, we applied ^(19)F solution NMR spectroscopy to monitor the conformational changes of G protein upon interact with detergent mimicking membrane and receptor. Our results show that there are two states equilibria in the G_(α)in apo states. The interaction of G_(α)with detergents will accelerate this conformational transformation and induce a state that tends to bind to GPCRs. Finally, the G_(α)proteins presented a fully activation state when they coupled to GPCRs.
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ERRATUM TO： PROTEIN CELL 2017, 8（2）：149-154 DOI 10.1007/S13238-016-0336-8In the original publication of this article the correspondence authors＇ e-mail has been missed out. The missing e-mail addresses are provided in this erratum.OPEN ACCESS This article is distributed under the terms of the Creative Commons Attribution 4.0 international License （http：//creativecommons.org/ licenses/by/4.0/）, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author（s） and the source, provide a link to the Creative Commons license, and indicate if changes were made.
