Cell-Free Synthesis of Selenoproteins in High Yield and Purity for Selective Protein Tagging

The selenol group of selenocysteine is much more nucleophilic than the thiol group of cysteine. Selenocysteine residues in proteins thus offer reactive points for rapid post-translational modification. Herein, we show that selenoproteins can be expressed in high yield and purity by cell-free protein synthesis by global substitution of cysteine by selenocysteine. Complete alkylation of solvent-exposed selenocysteine residues was achieved in 10 minutes with 4-chloromethylene dipicolinic acid (4Cl-MDPA) under conditions that left cysteine residues unchanged even after overnight incubation. Gd^III−Gd^III distances measured by double electron–electron resonance (DEER) experiments of maltose binding protein (MBP) containing two selenocysteine residues tagged with 4Cl-MDPA-Gd^III were indistinguishable from Gd^III−Gd^III distances measured of MBP containing cysteine reacted with 4Br-MDPA tags.     

王虔虔 双赢是硬道理

王虔虔在上任中建材(合肥)粉体科技装备有限公司(简称粉体公司)总经理之前,和分管部门的所有同事坐在同一间大办公室内。之所以和大家在同一个空间内工作,王虔虔是想更进一步拉近彼此之间的距离,以身作则潜移默化,传承公司文化,建立相互信任的团队。2012年,粉体公司决定向"提供系统解决方案的综合服务商"转变。     

Sensitivity analysis of Johnson-Cook material parameters on adiabatic shear in different directions

ABSTRACT

This study investigates the effects of strain, strain rates, and forming directions (RD-rolling direction, TD-transverse direction, and ND-normal direction) on adiabatic shear, via dynamic impact compression tests using the Split Hopkinson Pressure Bar (SHPB) apparatus. A modified Johnson-Cook (J-C) constitutive model is proposed, which used to analyse the influence of the constitutive parameters on the sensitivity of adiabatic shear, employing a finite element software. The different sensitivities of adiabatic shear under different directions are explained by combining microscopic analysis and results from mechanical responses. The results show that the sensitivity of adiabatic shear can be related to the time of stress collapse in the following trend: ND?>?TD?>?RD; the sensitivities of these constitutive parameters on adiabatic shear are calculated and compared.