title: Preparing semisynthetic and fully synthetic histones h3 and h4 to modify the nucleosome core.
abstract: The purpose of this chapter is to provide practical chemical ligation procedures to prepare histone proteins suitable for the reconstitution of nucleosomes with specific posttranslational modifications in the nucleosome core. Detailed methods are described for the efficient preparation of semisynthetic histones H3 and H4 with modifications near the C-terminus of the proteins by expressed protein ligation and desulfurization. Additionally, we present optimized protocols for solid phase peptide synthesis combined with sequential native chemical ligation to generate fully synthetic modified histone H3, here in the context of H3 lysine 56 acetylation (H3K56ac).
authors: Department of Biochemistry and The Ohio State Biochemistry Program, The Ohio State University, Columbus, OH, USA.
authors: John C
authors: JC
authors: Shimko
authors: Cecil J
authors: CJ
authors: Howard
authors: Michael G
authors: MG
authors: Poirier
authors: Jennifer J
authors: JJ
authors: Ottesen
mesh_terms: D000107:Acetylation; D001665:Binding Sites; D006657:Histones / Q000138:chemical synthesis* / Q000235:genetics; D008239:Lysine; D008967:Molecular Biology / Q000379:methods; D009707:Nucleosomes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational
chemical_list: D006657:Histones; D009707:Nucleosomes; D008239:Lysine
references: J Mol Biol. 2011 Apr 29;408(2):187-204
references: 21310161
references: Nucleic Acids Res. 2006 May 19;34(9):2653-62
references: 16714444
references: Chembiochem. 2011 Nov 4;12(16):2488-94
references: 21910203
references: Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10068-73
references: 10468563
references: Org Biomol Chem. 2006 Jun 7;4(11):2291-5
references: 16729139
references: Angew Chem Int Ed Engl. 2008;47(36):6851-5
references: 18651678
references: Proc Natl Acad Sci U S A. 2011 Aug 2;108(31):12711-6
references: 21768347
references: Nature. 1997 Sep 18;389(6648):251-60
references: 9305837
references: J Biol Chem. 2010 Apr 9;285(15):11045-50
references: 20147749
references: Science. 1974 May 24;184(4139):865-8
references: 4825888
references: Nucleic Acids Res. 2011 Aug;39(15):6465-74
references: 21576235
references: Curr Protoc Protein Sci. 2001 May;Chapter 18:Unit18.4
references: 18429140
references: Methods Mol Biol. 1999;119:1-16
references: 10804500
references: Angew Chem Int Ed Engl. 2007;46(48):9248-52
references: 18046687
references: Mol Cell. 2009 Dec 25;36(6):1086-94
references: 20064472
references: Int J Pept Protein Res. 1992 Sep-Oct;40(3-4):180-93
references: 1478777
references: J Biol Chem. 2009 Aug 28;284(35):23312-21
references: 19520870
references: J Org Chem. 1997 Jun 27;62(13):4307-4312
references: 11671751
references: Proc Natl Acad Sci U S A. 2006 Apr 25;103(17):6428-35
references: 16571659