Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligation
Summary: 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), recombinan...
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724006920 |
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| author | Yihang Xiao Kun Zou Mingxuan Wu |
| author_facet | Yihang Xiao Kun Zou Mingxuan Wu |
| author_sort | Yihang Xiao |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-87250744c89347999777a9baca6462e6 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-87250744c89347999777a9baca6462e62025-01-04T04:57:03ZengElsevierSTAR Protocols2666-16672025-03-0161103527Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligationYihang Xiao0Kun Zou1Mingxuan Wu2Department 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; Corresponding authorDepartment of Chemistry, School of Science, Westlake University, Hangzhou, Zhejiang Province 310030, ChinaDepartment 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; Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang Province 310024, China; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724006920Molecular BiologyProtein BiochemistryChemistry |
| spellingShingle | Yihang Xiao Kun Zou Mingxuan Wu Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligation STAR Protocols Molecular Biology Protein Biochemistry Chemistry |
| title | Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligation |
| title_full | Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligation |
| title_fullStr | Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligation |
| title_full_unstemmed | Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligation |
| title_short | Protocol for semisynthesis of histone H4 with site-specific modifications using irreversible sortase-mediated ligation |
| title_sort | protocol for semisynthesis of histone h4 with site specific modifications using irreversible sortase mediated ligation |
| topic | Molecular Biology Protein Biochemistry Chemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724006920 |
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