Peptide-Based Fluorescent Probes for Deacetylase and Decrotonylase Activity

Toward a General Platform for Real-Time Detection of Lysine Deacylation

Debra R. Rooker, Yuliya Klyubka, Ritika Gautam, Elisa Tomat, Daniela Buccella

Research output: Contribution to journalArticle

Abstract

Histone deacetylases regulate the acetylation levels of numerous proteins and play key roles in physiological processes and disease states. In addition to acetyl groups, deacetylases can remove other acyl modifications on lysines, the roles and regulation of which are far less understood. A peptide-based fluorescent probe for single-reagent, real-time detection of deacetylase activity that can be readily adapted for probing broader lysine deacylation, including decrotonylation, is reported. Following cleavage of the lysine modification, the probe undergoes rapid intramolecular imine formation that results in marked optical changes, thus enabling convenient detection of deacylase activity with good statistical Z′ factors for both absorption and fluorescence modalities. The peptide-based design offers broader isozyme scope than that of small-molecule analogues, and is suitable for probing both metal- and nicotinamide adenine dinucleotide (NAD+)-dependent deacetylases. With an effective sirtuin activity assay in hand, it is demonstrated that iron chelation by Sirtinol, a commonly employed sirtuin inhibitor, results in an enhancement in the inhibitory activity of the compound that may affect its performance in vivo.

Original languageEnglish (US)
Pages (from-to)496-504
Number of pages9
JournalChemBioChem
Volume19
Issue number5
DOIs
StatePublished - Mar 2 2018

Fingerprint

Fluorescent Dyes
Lysine
NAD
Peptides
Physiological Phenomena
Acetylation
Histone Deacetylases
Imines
Chelation
Isoenzymes
Assays
Iron
Hand
Fluorescence
Metals
Molecules
Proteins

Keywords

  • enzymes
  • fluorescence
  • peptides
  • probes
  • real-time detection

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

Cite this

Peptide-Based Fluorescent Probes for Deacetylase and Decrotonylase Activity : Toward a General Platform for Real-Time Detection of Lysine Deacylation. / Rooker, Debra R.; Klyubka, Yuliya; Gautam, Ritika; Tomat, Elisa; Buccella, Daniela.

In: ChemBioChem, Vol. 19, No. 5, 02.03.2018, p. 496-504.

Research output: Contribution to journalArticle

Rooker, Debra R. ; Klyubka, Yuliya ; Gautam, Ritika ; Tomat, Elisa ; Buccella, Daniela. / Peptide-Based Fluorescent Probes for Deacetylase and Decrotonylase Activity : Toward a General Platform for Real-Time Detection of Lysine Deacylation. In: ChemBioChem. 2018 ; Vol. 19, No. 5. pp. 496-504.
@article{4ea93fea026645cd8f8b431d0bc448fe,
title = "Peptide-Based Fluorescent Probes for Deacetylase and Decrotonylase Activity: Toward a General Platform for Real-Time Detection of Lysine Deacylation",
abstract = "Histone deacetylases regulate the acetylation levels of numerous proteins and play key roles in physiological processes and disease states. In addition to acetyl groups, deacetylases can remove other acyl modifications on lysines, the roles and regulation of which are far less understood. A peptide-based fluorescent probe for single-reagent, real-time detection of deacetylase activity that can be readily adapted for probing broader lysine deacylation, including decrotonylation, is reported. Following cleavage of the lysine modification, the probe undergoes rapid intramolecular imine formation that results in marked optical changes, thus enabling convenient detection of deacylase activity with good statistical Z′ factors for both absorption and fluorescence modalities. The peptide-based design offers broader isozyme scope than that of small-molecule analogues, and is suitable for probing both metal- and nicotinamide adenine dinucleotide (NAD+)-dependent deacetylases. With an effective sirtuin activity assay in hand, it is demonstrated that iron chelation by Sirtinol, a commonly employed sirtuin inhibitor, results in an enhancement in the inhibitory activity of the compound that may affect its performance in vivo.",
keywords = "enzymes, fluorescence, peptides, probes, real-time detection",
author = "Rooker, {Debra R.} and Yuliya Klyubka and Ritika Gautam and Elisa Tomat and Daniela Buccella",
year = "2018",
month = "3",
day = "2",
doi = "10.1002/cbic.201700582",
language = "English (US)",
volume = "19",
pages = "496--504",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "Wiley-VCH Verlag",
number = "5",

}

TY - JOUR

T1 - Peptide-Based Fluorescent Probes for Deacetylase and Decrotonylase Activity

T2 - Toward a General Platform for Real-Time Detection of Lysine Deacylation

AU - Rooker, Debra R.

AU - Klyubka, Yuliya

AU - Gautam, Ritika

AU - Tomat, Elisa

AU - Buccella, Daniela

PY - 2018/3/2

Y1 - 2018/3/2

N2 - Histone deacetylases regulate the acetylation levels of numerous proteins and play key roles in physiological processes and disease states. In addition to acetyl groups, deacetylases can remove other acyl modifications on lysines, the roles and regulation of which are far less understood. A peptide-based fluorescent probe for single-reagent, real-time detection of deacetylase activity that can be readily adapted for probing broader lysine deacylation, including decrotonylation, is reported. Following cleavage of the lysine modification, the probe undergoes rapid intramolecular imine formation that results in marked optical changes, thus enabling convenient detection of deacylase activity with good statistical Z′ factors for both absorption and fluorescence modalities. The peptide-based design offers broader isozyme scope than that of small-molecule analogues, and is suitable for probing both metal- and nicotinamide adenine dinucleotide (NAD+)-dependent deacetylases. With an effective sirtuin activity assay in hand, it is demonstrated that iron chelation by Sirtinol, a commonly employed sirtuin inhibitor, results in an enhancement in the inhibitory activity of the compound that may affect its performance in vivo.

AB - Histone deacetylases regulate the acetylation levels of numerous proteins and play key roles in physiological processes and disease states. In addition to acetyl groups, deacetylases can remove other acyl modifications on lysines, the roles and regulation of which are far less understood. A peptide-based fluorescent probe for single-reagent, real-time detection of deacetylase activity that can be readily adapted for probing broader lysine deacylation, including decrotonylation, is reported. Following cleavage of the lysine modification, the probe undergoes rapid intramolecular imine formation that results in marked optical changes, thus enabling convenient detection of deacylase activity with good statistical Z′ factors for both absorption and fluorescence modalities. The peptide-based design offers broader isozyme scope than that of small-molecule analogues, and is suitable for probing both metal- and nicotinamide adenine dinucleotide (NAD+)-dependent deacetylases. With an effective sirtuin activity assay in hand, it is demonstrated that iron chelation by Sirtinol, a commonly employed sirtuin inhibitor, results in an enhancement in the inhibitory activity of the compound that may affect its performance in vivo.

KW - enzymes

KW - fluorescence

KW - peptides

KW - probes

KW - real-time detection

UR - http://www.scopus.com/inward/record.url?scp=85041729858&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85041729858&partnerID=8YFLogxK

U2 - 10.1002/cbic.201700582

DO - 10.1002/cbic.201700582

M3 - Article

VL - 19

SP - 496

EP - 504

JO - ChemBioChem

JF - ChemBioChem

SN - 1439-4227

IS - 5

ER -