title: Synthesis of histone proteins by CPE ligation using a recombinant peptide as the C-terminal building block.
abstract: The post-translational modification of histones plays an important role in gene expression. We report herein on a method for synthesizing such modified histones by ligating chemically prepared N-terminal peptides and C-terminal recombinant peptide building blocks. Based on their chemical synthesis, core histones can be categorized as two types; histones H2A, H2B and H4 which contain no Cys residues, and histone H3 which contains a Cys residue(s) in the C-terminal region. A combination of native chemical ligation and desulphurization can be simply used to prepare histones without Cys residues. For the synthesis of histone H3, the endogenous Cys residue(s) must be selectively protected, while keeping the N-terminal Cys residue of the C-terminal building block that is introduced for purposes of chemical ligation unprotected. To this end, a phenacyl group was successfully utilized to protect endogenous Cys residue(s), and the recombinant peptide was ligated with a peptide containing a Cys-Pro ester (CPE) sequence as a thioester precursor. Using this approach it was possible to prepare all of the core histones H2A, H2B, H3 and H4 with any modifications. The resulting proteins could then be used to prepare a core histone library of proteins that have been post-translationally modified.
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan kawa@protein.osaka-u.ac.jp.
authors.forename: Toru
authors.initials: T
authors.lastname: Kawakami
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Yoshikawa
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Fujiyoshi
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Mishima
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan.
authors.forename: Hironobu
authors.initials: H
authors.lastname: Hojo
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan.
authors.forename: Shoji
authors.initials: S
authors.lastname: Tajima
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan; CREST, JST, Japan suetake@protein.osaka-u.ac.jp.
authors.forename: Isao
authors.initials: I
authors.lastname: Suetake
mesh_terms: D057205:Automation, Laboratory; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D020650:Combinatorial Chemistry Techniques; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004151:Dipeptides / Q000737:chemistry / Q000378:metabolism; D006657:Histones / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007564:Japan; D008239:Lysine / Q000737:chemistry / Q000378:metabolism; D008745:Methylation; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D060327:Solid-Phase Synthesis Techniques; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D004151:Dipeptides; D006657:Histones; D009842:Oligopeptides; D010446:Peptide Fragments; D011994:Recombinant Proteins; C028962:cysteinylproline; D008239:Lysine; D003545:Cysteine
keywords: CPE peptide; chemical ligation; histone; post-translational modification