title: Site-specific introduction of an acetyl-lysine mimic into peptides and proteins by cysteine alkylation.
abstract: Protein acetylation on Lys residues is recognized as a significant post-translational modification in cells, but it is often difficult to discern the direct structural and functional effects of individual acetylation events. Here we describe a new tool, methylthiocarbonyl-aziridine, to install acetyl-Lys mimics site-specifically into peptides and proteins by alkylation of Cys residues. We demonstrate that the resultant thiocarbamate modification can be recognized by the Brdt bromodomain and site-specific antiacetyl-Lys antibodies, is resistant to histone deacetylase cleavage, and can confer activation of the histone acetyltransferase Rtt109 by simulating autoacetylation. We also use this approach to obtain functional evidence that acetylation of CK2 protein kinase on Lys102 can stimulate its catalytic activity.
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Rong
authors.initials: R
authors.lastname: Huang
authors.forename: Marc A
authors.initials: MA
authors.lastname: Holbert
authors.forename: Mary Katherine
authors.initials: MK
authors.lastname: Tarrant
authors.forename: Sandrine
authors.initials: S
authors.lastname: Curtet
authors.forename: David R
authors.initials: DR
authors.lastname: Colquhoun
authors.forename: Beverley M
authors.initials: BM
authors.lastname: Dancy
authors.forename: Blair C
authors.initials: BC
authors.lastname: Dancy
authors.forename: Yousang
authors.initials: Y
authors.lastname: Hwang
authors.forename: Yong
authors.initials: Y
authors.lastname: Tang
authors.forename: Katrina
authors.initials: K
authors.lastname: Meeth
authors.forename: Ronen
authors.initials: R
authors.lastname: Marmorstein
authors.forename: Robert N
authors.initials: RN
authors.lastname: Cole
authors.forename: Saadi
authors.initials: S
authors.lastname: Khochbin
authors.forename: Philip A
authors.initials: PA
authors.lastname: Cole
mesh_terms: D000107:Acetylation; D000478:Alkylation; D000818:Animals; D001388:Aziridines / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D040761:Biomimetic Materials / Q000737:chemistry* / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism*; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D008239:Lysine / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D001388:Aziridines; D006657:Histones; D010455:Peptides; D011506:Proteins; C033132:aziridine; D008239:Lysine; D003545:Cysteine