Oral Presentation 11th Australian Peptide Conference 2015

Chemical synthesis of complex peptides and proteins by native chemical ligation (#41)

Thomas Durek 1
  1. University of Queensland, Brisbane, QLD, Australia

Secreted disulfide-rich polypeptides that modulate the activity if specific classes of membrane proteins are ubiquitous in nature and play key roles in various biological processes. They typically adopt highly ordered 3D structures and hold great promise as diagnostic tools and therapeutic agents. In the past, chemical synthesis has been instrumental for facilitating detailed SAR studies to fine-tune the properties of these molecules, however, this approach has been limited to polypeptides of less than about 40 amino acid residues. We adapted a robust and widely applicable strategy for gaining access to more complex molecules (50-100 AA) that is based on Kent’s native chemical ligation (NCL). In this presentation, I will cover some of our recent highlights including the synthesis of human anaphylatoxin C3a (77 AA), scorpion toxin OD1 (65 AA) and mambalgin-2 (57 AA) as well as some of our technological advances including the use of peptide-selenoesters instead of -thioesters in NCL. Strategies to extend the chemical ligation concept to even larger enzymes and proteins will be discussed, including the application of kinetically controlled ligation and expressed protein ligation.   

  1. Dawson, P. E., Muir, T. W., Clark-Lewis, I., and Kent, S. B. (1994) Synthesis of proteins by native chemical ligation, Science 266, 776-779.
  2. Schroeder, C. I., Rash, L. D., Vila-Farres, X., Rosengren, K. J., Mobli, M., King, G. F., Alewood, P. F., Craik, D. J., and Durek, T. (2014) Chemical Synthesis, 3D Structure, and ASIC Binding Site of the Toxin Mambalgin-2, Angew. Chem. Int. Ed. 53, 1017-1020
  3. Durek, T., and Alewood, P. F. (2011) Preformed Selenoesters Enable Rapid Native Chemical Ligation at Intractable Sites, Angew. Chem. Int. Ed. 50, 12042-12045.
  4. Durek, T., Torbeev, V. Y., and Kent, S. B. H. (2007) Convergent chemical synthesis and high-resolution x-ray structure of human lysozyme, Proc. Natl. Acad. Sci. U. S. A. 104, 4846-4851.