Novel artificial metalloenzymes by in vivo incorporation of metal-binding unnatural amino acids.

Novel artificial metalloenzymes by in vivo incorporation of metal-binding unnatural amino acids† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c4sc01525h Click here for additional data file.

Design of an enantioselective artificial metallo-hydratase enzyme containing an unnatural metal-binding amino acid.

The design of artificial metalloenzymes is a challenging, yet ultimately highly rewarding objective because of the potential for accessing new-to-nature reactions. One of the main challenges is identifying catalytically active substrate-metal cofactor-host geometries. The advent of expanded genetic code methods for the in vivo incorporation of non-canonical metal-binding amino acids into protei...

A kinetic safety gate controlling the delivery of unnatural amino acids to the ribosome.

Improving the yield of unnatural amino acid incorporation is an important challenge in producing novel designer proteins with unique chemical properties. Here we examine the mechanisms that restrict the incorporation of the fluorescent unnatural amino acid εNH2-Bodipy576/589-lysine (BOP-Lys) into a model protein. While the delivery of BOP-Lys-tRNA(Lys) to the ribosome is limited by its poor bin...

Steric and thermodynamic limits of design for the incorporation of large unnatural amino acids in aminoacyl-tRNA synthetase enzymes.

Orthogonal aminoacyl-tRNA synthetase/tRNA pairs from archaea have been evolved to facilitate site specific in vivo incorporation of unnatural amino acids into proteins in Escherichia coli. Using this approach, unnatural amino acids have been successfully incorporated with high translational efficiency and fidelity. In this study, CHARMM-based molecular docking and free energy calculations were ...

Metal-Ion-Coordinating Properties of Various Amino Acids, Investigation of the Essential Function in Biological Systems regarding to their Nano-Structure

The acidity constants of some amino acids (Am) were determined by potentiometric pH titration. The stability constants of the 1:1 complexes formed between M2+: Ca2+, Mg2+, Mn2+, Co2+, Ni2+, Cu2+ or Zn2+ and Am2-, were determined by potentiometric pH titration in aqueous solution (I = 0.1 M, NaNO3, 25°C). The order of the stability constants was reported. It is shown that the stability of the bi...