The Use of Nucleosome Substrates Improves Binding of SAM Analogs to SETD8.

Autor:	Strelow JM; Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, USA strelow_john_mark@lilly.com., Xiao M; Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, USA., Cavitt RN; Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, USA., Fite NC; Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, USA., Margolis BJ; Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, USA., Park KJ; Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, USA.
Jazyk:	angličtina
Zdroj:	Journal of biomolecular screening [J Biomol Screen] 2016 Sep; Vol. 21 (8), pp. 786-94. Date of Electronic Publication: 2016 Jul 01.
DOI:	10.1177/1087057116656596
Abstrakt:	SETD8 is the methyltransferase responsible for monomethylation of lysine at position 20 of the N-terminus of histone H4 (H4K20). This activity has been implicated in both DNA damage and cell cycle progression. Existing biochemical assays have utilized truncated enzymes containing the SET domain of SETD8 and peptide substrates. In this report, we present the development of a mechanistically balanced biochemical assay using full-length SETD8 and a recombinant nucleosome substrate. This improves the binding of SAM, SAH, and sinefungin by up to 10,000-fold. A small collection of inhibitors structurally related to SAM were screened and 40 compounds were identified that only inhibit SETD8 when a nucleosome substrate is used. (© 2016 Society for Laboratory Automation and Screening.)
Databáze:	MEDLINE
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