无细胞蛋白质合成(cell-free protein synthesis,CFPS)是一种在体外快速合成目标蛋白质的方法,通过构建含有CFPS系统的人造细胞,能够实现蛋白质的高通量表达和功能性膜蛋白的体外重构.本文详细综述了4种CFPS系统(包括大肠杆菌裂解液、...无细胞蛋白质合成(cell-free protein synthesis,CFPS)是一种在体外快速合成目标蛋白质的方法,通过构建含有CFPS系统的人造细胞,能够实现蛋白质的高通量表达和功能性膜蛋白的体外重构.本文详细综述了4种CFPS系统(包括大肠杆菌裂解液、兔网织红细胞裂解液、小麦胚芽提取物、酵母提取物)的适用范围和优缺点,总结了基于CFPS系统构建的人造细胞体系内蛋白质合成的研究现状,以及该领域面临的挑战及未来的发展方向.展开更多
An implicit solvent coarse-grained （CG） lipid model using three beads to reflect the basically molecular structure of two-tailed lipid is developed. In this model, the nonbonded interaction employs a variant MIE pot...An implicit solvent coarse-grained （CG） lipid model using three beads to reflect the basically molecular structure of two-tailed lipid is developed. In this model, the nonbonded interaction employs a variant MIE potential and the bonded interaction utilizes a Harmonic potential form. The CG force field parameters are achieved by matching the structural and mechan-ical properties of dipalmitoylphosphatidylcholine （DPPC） bilayers. The model successfully reproduces the formation of lipid bilayer from a random initial state and the spontaneous vesiculation of lipid bilayer from a disk-like structure. After that, the model is used to sys-tematically study the vesiculation processes of spherical and cylindrical lipid droplets. The results show that the present CG model can effectively simulate the formation and evolution of mesoscale complex vesicles.展开更多
Lysozyme, myoglobin and BSA were used as models of globular proteins covering a wide range of pl. The purpose is to extend the studies to anionic lipid bilayers. Electrostatics is studied in cationic protein adsorptio...Lysozyme, myoglobin and BSA were used as models of globular proteins covering a wide range of pl. The purpose is to extend the studies to anionic lipid bilayers. Electrostatics is studied in cationic protein adsorption to zwitterionic PC and anionic mixed PC/PG SUVs. Protein adsorption is investigated in SUVs along with changes of fluorescence emission spectra. Partition coefficients and cooperativity parameters are calculated. At pl binding obtains maximum while at lower or higher pHs binding decreases. In Gouy-Chapman formalism activity coefficient goes with square charge, which deviations indicate asymmetric location of anionic phospholipid in the inner leaflet, in mixed SUVs for lysozyme- and myoglobin-PC/PG systems, in agreement with experiments and molecular dynamics simulations. Vesicles bind myoglobin anti-cooperatively while lysozyme-BSA cooperativitivey. A model is proposed for both, which composes two protein sub-layers with different structures and properties. Hill coefficient reflects subunit cooperativity of bi and tridomain proteins.展开更多
文摘无细胞蛋白质合成(cell-free protein synthesis,CFPS)是一种在体外快速合成目标蛋白质的方法,通过构建含有CFPS系统的人造细胞,能够实现蛋白质的高通量表达和功能性膜蛋白的体外重构.本文详细综述了4种CFPS系统(包括大肠杆菌裂解液、兔网织红细胞裂解液、小麦胚芽提取物、酵母提取物)的适用范围和优缺点,总结了基于CFPS系统构建的人造细胞体系内蛋白质合成的研究现状,以及该领域面临的挑战及未来的发展方向.
基金We thank Professor Dr. Markus Deserno （Department of Physics, Carnegie Mellon University）, Jemal Guven （Institute of Nuclear Science, Universidad Nacional Autonoma de Mexico）, and Zhan-chun Tu （Department of Physics, Beijing Normal University） for their valuable advice. This work is supported by the National Natural Science Foundation of China （No.20974078, No.21274107, and No.91127046）. The computation and simulation are partly carried out in High Performance Computing Center of Tianjin University.
文摘An implicit solvent coarse-grained （CG） lipid model using three beads to reflect the basically molecular structure of two-tailed lipid is developed. In this model, the nonbonded interaction employs a variant MIE potential and the bonded interaction utilizes a Harmonic potential form. The CG force field parameters are achieved by matching the structural and mechan-ical properties of dipalmitoylphosphatidylcholine （DPPC） bilayers. The model successfully reproduces the formation of lipid bilayer from a random initial state and the spontaneous vesiculation of lipid bilayer from a disk-like structure. After that, the model is used to sys-tematically study the vesiculation processes of spherical and cylindrical lipid droplets. The results show that the present CG model can effectively simulate the formation and evolution of mesoscale complex vesicles.
文摘Lysozyme, myoglobin and BSA were used as models of globular proteins covering a wide range of pl. The purpose is to extend the studies to anionic lipid bilayers. Electrostatics is studied in cationic protein adsorption to zwitterionic PC and anionic mixed PC/PG SUVs. Protein adsorption is investigated in SUVs along with changes of fluorescence emission spectra. Partition coefficients and cooperativity parameters are calculated. At pl binding obtains maximum while at lower or higher pHs binding decreases. In Gouy-Chapman formalism activity coefficient goes with square charge, which deviations indicate asymmetric location of anionic phospholipid in the inner leaflet, in mixed SUVs for lysozyme- and myoglobin-PC/PG systems, in agreement with experiments and molecular dynamics simulations. Vesicles bind myoglobin anti-cooperatively while lysozyme-BSA cooperativitivey. A model is proposed for both, which composes two protein sub-layers with different structures and properties. Hill coefficient reflects subunit cooperativity of bi and tridomain proteins.
