Poster abstracts
Poster number 5 submitted by Ryan Yu
Hybrid-Phase Ligation and Convergent Solid-Phase Ligation for Efficient Protein Total Synthesis
Ryan Yu (OSBP), Ziyong Hong (Chemistry)
Abstract:
Protein total synthesis is a powerful technique that allows the preparation of chemically diverse proteins not possible using conventional methods. Here, we develop hybrid solid-solution phase native chemical ligation that combines the efficiency of ligation on solid-phase with the yield of ligation in solution-phase. We demonstrate the effectiveness of this method through the synthesis of histone H4 and CENP-A proteins from four and five peptide segments.
Conventional convergent ligation approaches reach an effective size barrier through the chemical limits of solid phase peptide synthesis combined with practical limits of multiple solution ligation/purification steps. We demonstrate here preliminary results in convergent hybrid phase ligation, which has the potential to overcome these limits. Here, short peptide segments can be ligated on the solid phase to generate large ligation-compatible segments not accessible through direct synthesis. These large segments can be released from resin and ligated in solution to produce the full-length protein. We describe preliminary results in developing ligation handles compatible with this approach in the context of CENP-A and the 233-residue linker histone H1.2.
Keywords: peptide synthesis, post-translational modifications, histone