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.     

A Quest for (sila)0-4cyclopentasilylenes and their Arduengo Analogs by DFT

Silicon - Following our interest in exotic silylenes, here we compare and contrast 20 novel five-membered cyclic silylenes, including saturated (sila)0–4 cyclopentasilylenes (1–10) and...     

Steels for Bridge Structures

Metallurgist - We consider domestic and foreign standards for rolled metals used in bridge building. Domestic standards contain elevated requirements to the reliability of rolled metals in terms of...