A chemical method for labeling lysine methyltransferase substrates.
Abstract
Several protein lysine methyltransferases (PKMTs) modify histones to regulate chromatin-dependent
cellular processes, such as transcription, DNA replication and DNA damage repair.
PKMTs are likely to have many additional substrates in addition to histones, but relatively
few nonhistone substrates have been characterized, and the substrate specificity for
many PKMTs has yet to be defined. Thus, new unbiased methods are needed to find PKMT
substrates. Here, we describe a chemical biology approach for unbiased, proteome-wide
identification of novel PKMT substrates. Our strategy makes use of an alkyne-bearing
S-adenosylmethionine (SAM) analogue, which is accepted by the PKMT, SETDB1, as a cofactor,
resulting in the enzymatic attachment of a terminal alkyne to its substrate. Such
labeled proteins can then be treated with azide-functionalized probes to ligate affinity
handles or fluorophores to the PKMT substrates. As a proof-of-concept, we have used
SETDB1 to transfer the alkyne moiety from the SAM analogue onto a recombinant histone
H3 substrate. We anticipate that this chemical method will find broad use in epigenetics
to enable unbiased searches for new PKMT substrates by using recombinant enzymes and
unnatural SAM cofactors to label and purify many substrates simultaneously from complex
organelle or cell extracts.
Type
Journal articleSubject
AlkynesMethyltransferases
Lysine
S-Adenosylmethionine
Recombinant Proteins
Staining and Labeling
Molecular Structure
Substrate Specificity
Epigenomics
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https://hdl.handle.net/10161/19701Published Version (Please cite this version)
10.1002/cbic.201000433Publication Info
Binda, Olivier; Boyce, Michael; Rush, Jason S; Palaniappan, Krishnan K; Bertozzi,
Carolyn R; & Gozani, Or (2011). A chemical method for labeling lysine methyltransferase substrates. Chembiochem : a European journal of chemical biology, 12(2). pp. 330-334. 10.1002/cbic.201000433. Retrieved from https://hdl.handle.net/10161/19701.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Michael Scott Boyce
Associate Professor of Biochemistry
The Boyce Lab studies mammalian cell signaling through protein glycosylation. For
the latest news, project information and publications from our group, please visit
our web site at http://www.boycelab.org or follow us on Twitter at https://twitter.com/BoyceLab.

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