Convergent Hybrid-phase Ligation Synthetic Strategy Combined with Direct Refolding Method to Improve Synthetic Yield of Centromeric Protein A
Author | : Xiaoyu Zhang |
Publisher | : |
Total Pages | : |
Release | : 2020 |
ISBN-10 | : OCLC:1322111968 |
ISBN-13 | : |
Rating | : 4/5 (68 Downloads) |
Book excerpt: Histone proteins are a series of post-translational modification (PTM) heavy proteins. Total protein chemical synthesis possesses advantages in preparation of these protein samples containing specific PTM(s). However, the synthetic yield is always one of the biggest limitation of total protein chemical synthesis, especially when preparing large protein sample. Our lab had developed a synthetic strategy to total synthesize the H3 variant, Centromeric protein A (Cenp- A). Even though the synthesis was successful, the overall synthetic yield still did not fulfil our expectation. Here, we have developed a new convergent hybrid-phase synthetic strategy combined with direct histone octamer refolding and applied on the total synthesis of Cenp-A. This new strategy could maximally utilize the advantage of solid-phase native chemical ligation and avoid the any solution-phase purification steps which turned out to be the main reason of causing yield loss during the synthesis. This new synthetic strategy holds potential of greatly improving the synthetic yield of Cenp-A and being applied on the total synthesis of other large proteins.