A fluorescence resonance energy transfer-based method for histone methyltransferases

Kanchan Devkota; Brian Lohse; Camilla Nyby Jakobsen; Jens Berthelsen; Rasmus Prætorius Clausen

doi:10.1016/j.ab.2015.02.012

A fluorescence resonance energy transfer-based method for histone methyltransferases

Kanchan Devkota, Brian Lohse, Camilla Nyby Jakobsen, Jens Berthelsen, Rasmus Prætorius Clausen

1 Citation (Scopus)

Abstract

Abstract A simple dye-quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye and a quencher, respectively. When lysine-9 residues in the peptides were methylated, they were protected from cleavage by endoproteinase-EndoLysC, whereas unmethylated peptides were cleaved, resulting in an increase in fluorescent intensity.

Original language	English
Journal	Analytical Biochemistry
Volume	476
Pages (from-to)	78-80
Number of pages	3
ISSN	0003-2697
DOIs	https://doi.org/10.1016/j.ab.2015.02.012
Publication status	Published - 1 May 2015

Keywords

Faculty of Health and Medical Sciences

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title = "A fluorescence resonance energy transfer-based method for histone methyltransferases",

abstract = "Abstract A simple dye-quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye and a quencher, respectively. When lysine-9 residues in the peptides were methylated, they were protected from cleavage by endoproteinase-EndoLysC, whereas unmethylated peptides were cleaved, resulting in an increase in fluorescent intensity.",

keywords = "Faculty of Health and Medical Sciences",

author = "Kanchan Devkota and Brian Lohse and {Nyby Jakobsen}, Camilla and Jens Berthelsen and Clausen, {Rasmus Pr{\ae}torius}",

year = "2015",

month = may,

day = "1",

doi = "10.1016/j.ab.2015.02.012",

language = "English",

volume = "476",

pages = "78--80",

journal = "Analytical Biochemistry",

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T1 - A fluorescence resonance energy transfer-based method for histone methyltransferases

AU - Devkota, Kanchan

AU - Lohse, Brian

AU - Nyby Jakobsen, Camilla

AU - Berthelsen, Jens

AU - Clausen, Rasmus Prætorius

PY - 2015/5/1

Y1 - 2015/5/1

N2 - Abstract A simple dye-quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye and a quencher, respectively. When lysine-9 residues in the peptides were methylated, they were protected from cleavage by endoproteinase-EndoLysC, whereas unmethylated peptides were cleaved, resulting in an increase in fluorescent intensity.

AB - Abstract A simple dye-quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye and a quencher, respectively. When lysine-9 residues in the peptides were methylated, they were protected from cleavage by endoproteinase-EndoLysC, whereas unmethylated peptides were cleaved, resulting in an increase in fluorescent intensity.

KW - Faculty of Health and Medical Sciences

U2 - 10.1016/j.ab.2015.02.012

DO - 10.1016/j.ab.2015.02.012

M3 - Journal article

C2 - 25703602

SN - 0003-2697

VL - 476

SP - 78

EP - 80

JO - Analytical Biochemistry

JF - Analytical Biochemistry

ER -

A fluorescence resonance energy transfer-based method for histone methyltransferases

Abstract

Keywords

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