Utilizing intein trans-splicing for in vivo generation of site-specifically modified proteins Journal Article
| Authors: | Maksimovic, I.; Ray, D.; Zheng, Q.; David, Y. |
| Article Title: | Utilizing intein trans-splicing for in vivo generation of site-specifically modified proteins |
| Abstract: | Many cellular processes as well as their associated pathologies are regulated by protein post-translational modifications (PTMs). Understanding the precise roles of these adducts hinges on the development of methods to robustly and site-specifically manipulate proteins in their physiological environments. Recently, ultrafast intein protein trans-splicing (PTS) was harnessed to incorporate site-specific modifications on cellular chromatin in live cells. In this chapter, we present the protocols for the generation of synthetic modifications on native chromatin as well as highlight the capabilities of this methodology. © 2019 Elsevier Inc. |
| Keywords: | controlled study; protein expression; human cell; nonhuman; confocal microscopy; amino acid sequence; genetic transfection; protein purification; protein synthesis; epigenetics; western blotting; histone h2b; endocytosis; peptide synthesis; protein modification; chromatin structure; conjugation; solid phase synthesis; histone modification; electrospray mass spectrometry; reversed phase high performance liquid chromatography; chemical biology; trans splicing; protein engineering; disulfide bond; intein; cell penetrating peptide; chromatin remodeling; human; live cell imaging; time-lapse microscopy; hek293t cell line; protein splicing; ultrafast split inteins |
| Journal Title: | Methods in Enzymology |
| Volume: | 626 |
| ISSN: | 0076-6879 |
| Publisher: | Academic Press |
| Date Published: | 2019-01-01 |
| Start Page: | 203 |
| End Page: | 222 |
| Language: | English |
| DOI: | 10.1016/bs.mie.2019.07.015 |
| PUBMED: | 31606075 |
| PROVIDER: | scopus |
| PMCID: | PMC8080263 |
| DOI/URL: | |
| Notes: | Chapter 9 in "Post-translational Modifications That Modulate Enzyme Activity" (ISBN: 978-0-12-818669-5) -- Source: Scopus |
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