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Updated: Jun 4, 2025
Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues
Published on: November 22, 2014
Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated
Yihang Xiao1, Kun Zou2, Mingxuan Wu3
1Department of Chemistry, School of Science, Westlake University, Hangzhou, Zhejiang Province 310030, China; Institute of Natural Sciences, Westlake Institute for Advanced Study, Hangzhou, Zhejiang Province 310024, China.
None:
Post-translational modifications (PTMs) of histone H4 play significant roles in the regulation of chromatin status. Here, we present a protocol for semisynthesis of histone H4 by sortase-mediated ligation (SML). We describe steps for solid-phase peptide synthesis of H4R40C(1-42), recombinant expression and purification of H4(41-102), expression and purification of eSrt(2A-9), and preparation of acrylamidine. We then detail procedures for SML of histone H4. This protocol can also be applied to the preparation of homogenous proteins with PTMs. For complete details on the use and execution of this protocol, please refer to Xiao et al.1,2.
