-
Something wrong with this record ?
Continuous Sirtuin/HDAC (histone deacetylase) activity assay using thioamides as PET (Photoinduced Electron Transfer)-based fluorescence quencher
M. Zessin, M. Meleshin, Z. Simic, D. Kalbas, M. Arbach, P. Gebhardt, J. Melesina, S. Liebscher, F. Bordusa, W. Sippl, C. Barinka, M. Schutkowski
Bioorganic chemistry.
2021 ;
117 (-) :
105425.
[pub] 20211012
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
Persistent link
https://www.medvik.cz/link/bmc22011776
- MeSH
- Fluorescent Dyes chemistry pharmacology MeSH
- Photochemical Processes MeSH
- Histone Deacetylases chemistry genetics metabolism MeSH
- Humans MeSH
- Molecular Structure MeSH
- Positron-Emission Tomography * MeSH
- Sirtuins antagonists & inhibitors chemistry metabolism MeSH
- Thioamides chemistry pharmacology MeSH
- Electron Transport MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Histone deacylase 11 and human sirtuins are able to remove fatty acid-derived acyl moieties from the ε-amino group of lysine residues. Specific substrates are needed for investigating the biological functions of these enzymes. Additionally, appropriate screening systems are required for identification of modulators of enzymatic activities of HDAC11 and sirtuins. We designed and synthesized a set of activity probes by incorporation of a thioamide quencher unit into the fatty acid-derived acyl chain and a fluorophore in the peptide sequence. Systematic variation of both fluorophore and quencher position resulted "super-substrates" with catalytic constants of up to 15,000,000 M-1s-1 for human sirtuin 2 (Sirt2) enabling measurements using enzyme concentrations down to 100 pM in microtiter plate-based screening formats. It could be demonstrated that the stalled intermediate formed by the reaction of Sirt2-bound thiomyristoylated peptide and NAD+ has IC50 values below 200 pM.
References provided by Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc22011776
- 003
- CZ-PrNML
- 005
- 20220506125816.0
- 007
- ta
- 008
- 220425s2021 xxu f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1016/j.bioorg.2021.105425 $2 doi
- 035 __
- $a (PubMed)34695733
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a xxu
- 100 1_
- $a Zessin, Matthes $u Department of Medicinal Chemistry, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 245 10
- $a Continuous Sirtuin/HDAC (histone deacetylase) activity assay using thioamides as PET (Photoinduced Electron Transfer)-based fluorescence quencher / $c M. Zessin, M. Meleshin, Z. Simic, D. Kalbas, M. Arbach, P. Gebhardt, J. Melesina, S. Liebscher, F. Bordusa, W. Sippl, C. Barinka, M. Schutkowski
- 520 9_
- $a Histone deacylase 11 and human sirtuins are able to remove fatty acid-derived acyl moieties from the ε-amino group of lysine residues. Specific substrates are needed for investigating the biological functions of these enzymes. Additionally, appropriate screening systems are required for identification of modulators of enzymatic activities of HDAC11 and sirtuins. We designed and synthesized a set of activity probes by incorporation of a thioamide quencher unit into the fatty acid-derived acyl chain and a fluorophore in the peptide sequence. Systematic variation of both fluorophore and quencher position resulted "super-substrates" with catalytic constants of up to 15,000,000 M-1s-1 for human sirtuin 2 (Sirt2) enabling measurements using enzyme concentrations down to 100 pM in microtiter plate-based screening formats. It could be demonstrated that the stalled intermediate formed by the reaction of Sirt2-bound thiomyristoylated peptide and NAD+ has IC50 values below 200 pM.
- 650 _2
- $a transport elektronů $7 D004579
- 650 _2
- $a fluorescenční barviva $x chemie $x farmakologie $7 D005456
- 650 _2
- $a histondeacetylasy $x chemie $x genetika $x metabolismus $7 D006655
- 650 _2
- $a lidé $7 D006801
- 650 _2
- $a molekulární struktura $7 D015394
- 650 _2
- $a fotochemické procesy $7 D055668
- 650 12
- $a pozitronová emisní tomografie $7 D049268
- 650 _2
- $a sirtuiny $x antagonisté a inhibitory $x chemie $x metabolismus $7 D037761
- 650 _2
- $a thioamidy $x chemie $x farmakologie $7 D013854
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Meleshin, Marat $u Department of Enzymology, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Simic, Zeljko $u Department of Enzymology, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Kalbas, Diana $u Department of Enzymology, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Arbach, Miriam $u Department of Enzymology, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Gebhardt, Philip $u Department of Enzymology, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Melesina, Jelena $u Department of Medicinal Chemistry, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Liebscher, Sandra $u Department of Natural Product Biochemistry, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Bordusa, Frank $u Department of Natural Product Biochemistry, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Sippl, Wolfgang $u Department of Medicinal Chemistry, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
- 700 1_
- $a Barinka, Cyril $u Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic
- 700 1_
- $a Schutkowski, Mike $u Department of Enzymology, Charles Tanford Protein Center, Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany. Electronic address: mike.schutkowski@biochemtech.uni-halle.de
- 773 0_
- $w MED00000771 $t Bioorganic chemistry $x 1090-2120 $g Roč. 117, č. - (2021), s. 105425
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/34695733 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y p $z 0
- 990 __
- $a 20220425 $b ABA008
- 991 __
- $a 20220506125808 $b ABA008
- 999 __
- $a ok $b bmc $g 1789396 $s 1162974
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2021 $b 117 $c - $d 105425 $e 20211012 $i 1090-2120 $m Bioorganic chemistry $n Bioorg Chem $x MED00000771
- LZP __
- $a Pubmed-20220425
