Protein Engineering with Noncoded Amino Acids: Applications to Hirudin

Abstract

The incorporation of noncoded amino acids into proteins has emerged as a novel and promising approach in protein science. Several different strategies are being pursued to incorporate noncoded amino acids, including peptide synthesis native chemical ligation, enzyme‐catalyzed semisynthesis, biosynthetic incorporation via auxotrophic bacterial strain expression, and nonsense suppression methodologies in cell‐free or whole‐cell expression systems. This article discusses the use of noncoded amino acids in structure‐activity relationship (SAR) studies of hirudin binding to thrombin, as well as the incorporation of noncoded analogs of tryptophan and tyrosine as spectroscopic probes for studying the hirudin‐thrombin interaction.