title: Genetically encoding lysine modifications on histone H4.
abstract: Post-translational modifications of proteins are important modulators of protein function. In order to identify the specific consequences of individual modifications, general methods are required for homogeneous production of modified proteins. The direct installation of modified amino acids by genetic code expansion facilitates the production of such proteins independent of the knowledge and availability of the enzymes naturally responsible for the modification. The production of recombinant histone H4 with genetically encoded modifications has proven notoriously difficult in the past. Here, we present a general strategy to produce histone H4 with acetylation, propionylation, butyrylation, and crotonylation on lysine residues. We produce homogeneous histone H4 containing up to four simultaneous acetylations to analyze the impact of the modifications on chromatin array compaction. Furthermore, we explore the ability of antibodies to discriminate between alternative lysine acylations by incorporating these modifications in recombinant histone H4.
authors.affiliation: †Free Floater (Junior) Research Group "Applied Synthetic Biology", Institute for Microbiology and Genetics, Georg-August University Göttingen, Justus-von-Liebig Weg 11, 37077 Göttingen, Germany.
authors.forename: Bryan J
authors.initials: BJ
authors.lastname: Wilkins
authors.affiliation: †Free Floater (Junior) Research Group "Applied Synthetic Biology", Institute for Microbiology and Genetics, Georg-August University Göttingen, Justus-von-Liebig Weg 11, 37077 Göttingen, Germany.
authors.forename: Liljan E
authors.initials: LE
authors.lastname: Hahn
authors.affiliation: †Free Floater (Junior) Research Group "Applied Synthetic Biology", Institute for Microbiology and Genetics, Georg-August University Göttingen, Justus-von-Liebig Weg 11, 37077 Göttingen, Germany.
authors.forename: Svenja
authors.initials: S
authors.lastname: Heitmüller
authors.affiliation: ‡Institute for Organic and Biomolecular Chemistry, Georg-August University Göttingen, Tammannstrasse 2, 37077 Göttingen, Germany.
authors.forename: Holm
authors.initials: H
authors.lastname: Frauendorf
authors.affiliation: §Institute for Microbiology and Genetics, Georg-August University Göttingen, Grisebachstrasse 8, 37077 Göttingen, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Valerius
authors.affiliation: §Institute for Microbiology and Genetics, Georg-August University Göttingen, Grisebachstrasse 8, 37077 Göttingen, Germany.
authors.forename: Gerhard H
authors.initials: GH
authors.lastname: Braus
authors.affiliation: †Free Floater (Junior) Research Group "Applied Synthetic Biology", Institute for Microbiology and Genetics, Georg-August University Göttingen, Justus-von-Liebig Weg 11, 37077 Göttingen, Germany.
authors.forename: Heinz
authors.initials: H
authors.lastname: Neumann
mesh_terms: D000107:Acetylation; D002843:Chromatin / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D009707:Nucleosomes; D015202:Protein Engineering / Q000379:methods*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D002843:Chromatin; D029721:Drosophila Proteins; D006657:Histones; D009707:Nucleosomes; D011993:Recombinant Fusion Proteins; D010450:Endopeptidases; C099488:TEV protease; D008239:Lysine