Rxivist logo

Convergent Hybrid Phase Ligation Strategy for Efficient Total Synthesis of Large Proteins Demonstrated for 212-residue Linker Histone H1.2

By Ziyong Z Hong, Ruixuan R. Yu, Xiaoyu Zhang, Allison M Webb, Nathaniel L Burge, Michael G Poirier, Jennifer J. Ottesen

Posted 06 Jun 2019
bioRxiv DOI: 10.1101/661744

Simple and efficient total chemical synthesis of large proteins remains a significant challenge. Here, we report development of a convergent hybrid phase native chemical ligation (CHP-NCL) strategy that should be generally applicable for facile preparation of large proteins. Key to the strategy is the use of sequential ligation on the solid phase for the directed assembly of ~100-residue segments from short, synthetically accessible peptide components. These segments can then be assembled via convergent solution phase ligation, exploiting o-aminoaniline as a chemically flexible cryptic thioester with multiple activation modalitiies on resin and in situ. We demonstrate the feasibility of our approach through the total synthesis of 212-residue linker histone H1.2 in unmodified, phosphorylated, and citrullinated forms, each from eight component peptide segments. We further demonstrate that fully synthetic H1.2 replicates the binding interactions of linker histones to intact mononucleosomes, as a proxy for the essential function of linker histones in the formation and regulation of higher order chromatin structure.

Download data

  • Downloaded 458 times
  • Download rankings, all-time:
    • Site-wide: 54,941
    • In biochemistry: 1,444
  • Year to date:
    • Site-wide: 59,463
  • Since beginning of last month:
    • Site-wide: 45,309

Altmetric data


Downloads over time

Distribution of downloads per paper, site-wide


PanLingua

Sign up for the Rxivist weekly newsletter! (Click here for more details.)


News