Protein Mutagenesis using Unnatural amino acids by Frances Valiyaveetil (Oregon Health and Science)

Submitted by Patricia B. O'Hara on Monday, 3/7/2011, at 10:54 PM

1. Peptide synthesis (has a size limitation of 60 aa)

2. Semi-synthesis (make three 60-mers and add them together either chemically or biochemically

3. Biologically:

in vitro protein synthesis: use a nonsense suppressor, add in a rare stop codon with designed tRNA complement with you acylated unnatural amino acid, off you go

in vivo protein synthesis:  Peter Schultz has a bunch of modified tRNA synthetases that can make unnatural tRNAs and assemble them in vivo.  Limitations are that all unnatural amino acids (about 100 of them) are Tyr derivatives w tRNA’s to match.