Site-specific introduction of an acetyl-lysine mimic into peptides and proteins by cysteine alkylation.
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.
Huang
Rong
R
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Holbert
Marc A
MA
Tarrant
Mary Katherine
MK
Curtet
Sandrine
S
Colquhoun
David R
DR
Dancy
Beverley M
BM
Dancy
Blair C
BC
Hwang
Yousang
Y
Tang
Yong
Y
Meeth
Katrina
K
Marmorstein
Ronen
R
Cole
Robert N
RN
Khochbin
Saadi
S
Cole
Philip A
PA
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
D001388:Aziridines; D006657:Histones; D010455:Peptides; D011506:Proteins; C033132:aziridine; D008239:Lysine; D003545:Cysteine