Fluorescence-labeled octapeptides as substrates for histone deacetylase

Citation
K. Hoffmann et al., Fluorescence-labeled octapeptides as substrates for histone deacetylase, BIOCONJ CHE, 12(1), 2001, pp. 51-55
Citations number
21
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Chemistry & Analysis
Journal title
BIOCONJUGATE CHEMISTRY
ISSN journal
1043-1802 → ACNP
Volume
12
Issue
1
Year of publication
2001
Pages
51 - 55
Database
ISI
SICI code
1043-1802(200101/02)12:1<51:FOASFH>2.0.ZU;2-D
Abstract
The determination of histone deacetylase (HDAC) activity and the screening of potential inhibitors is gaining increasing importance due to the involve ment of HDAC in transcription regulation. The level of histone acetylation can be modulated by HDAC inhibitors resulting in differentiation and/or apo ptosis in cancer cells. We have previously reported the development of a no nisotopic assay for HDAC using a fluorescent derivative of epsilon -acetyl lysine. Here we report fluorescein-labeled octapeptides which are substrate s for HDAC that bear closer resemblance to the native substrate. HPLC with fluorescence detection is successfully applied to the analysis of the time- and site-dependent deacetylation. LC-MS analyses are used to confirm the f indings. The observed selectivity toward one of two possible deacetylation sites might result from steric hindrance by the label but the methodology p resented here could be applied to similar larger peptides which might be im proved tools to characterize HDAC site selectivity in vitro.