Effects of Non-Natural Amino Acid Incorporation into the Enzyme Core Region on Enzyme Structure and Function.

摘要: Techniques to incorporate non-natural amino acids (NNAAs) have enabled biosynthesis of proteins containing new building blocks with unique structures, chemistry, and reactivity that are not found in natural acids. It is crucial understand how incorporation NNAAs affects protein function because NNAA may perturb critical a target protein. This study investigates the site-specific catalytic properties an enzyme. A hydrophobic bulky sidechain, 3-(2-naphthyl)-alanine (2Nal), was site-specifically incorporated at six different positions core model enzyme, murine dihydrofolate reductase (mDHFR). The mDHFR variants greater change van der Waals volume upon 2Nal exhibited reduction efficiency. Similarly, steric incompatibility calculated using RosettaDesign, stability calculation program, correlated changes