Factors affecting the substrate specificity of histone deacetylases.
Histone deacetylases (HDACs) catalyze the deacetylation of epsilon-acetyl-lysine residues within the N-terminal tail of core histones and thereby mediate changes in the chromatin structure and regulate gene expression in eukaryotic cells. So far, surprisingly little is known about the substrate specificities of different HDACs. Here, we prepared a library of fluorogenic tripeptidic substrates of the general format Ac-P(-2)-P(-1)-Lys(Ac)-MCA (P(-1), P(-2)=all amino acids except cysteine) and measured their HDAC-dependent conversion in a standard fluorogenic HDAC assay. Different HDAC subtypes can be ranked according to their substrate selectivity: HDAH > HDAC8 > HDAC1 > HDAC3 > HDAC6. HDAC1, HDAC3, and HDAC6 exhibit a similar specificity profile, whereas both HDAC8 and HDAH have rather distinct profiles. Furthermore, it was shown that second-site modification (e.g., phosphorylation) of substrate sequences as well as corepressor binding can modulate the selectivity of enzymatic substrate conversion.
Riester
Daniel
D
Department of Molecular Genetics and Preparative Molecular Biology, Institute for Microbiology and Genetics, University of Goettingen, Grisebachstr. 8, 37077 Goettingen, Germany.
Hildmann
Christian
C
Grünewald
Sylvia
S
Beckers
Thomas
T
Schwienhorst
Andreas
A
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001665:Binding Sites; D004789:Enzyme Activation; D006655:Histone Deacetylases / Q000737:chemistry* / Q000145:classification*; D008969:Molecular Sequence Data; D011485:Protein Binding; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D006655:Histone Deacetylases