Profiling deacetylase activities in cell lysates with peptide arrays and SAMDI mass spectrometry

Hsin Yu Kuo, Teresa A. Deluca, William M Miller, Milan Mrksich

Research output: Contribution to journalReview article

22 Citations (Scopus)

Abstract

The development of arrays that can profile molecular activities in cells is important to understanding signaling pathways in normal and pathological settings. While oligonucleotide arrays are now routinely used to profile global gene expression, there is still a lack of tools for profiling enzyme activities in cell lysates. This paper describes the combination of peptide arrays formed on self-assembled monolayers and mass spectrometry to provide a label-free approach for identifying patterns of enzyme activities in cell lysates. The approach is demonstrated by profiling lysine deacetylase (KDAC) activities in cell lysates of the CHRF megakaryocytic (Mk) cell line. Class-specific deacetylase inhibitors were used to show that terminal Mk differentiation of CHRF cells is marked by a pronounced decrease in sirtuin activity and by little change in activity of KDACs 1-11. This work establishes a platform that can be used to identify changes in global activity profiles of cell lysates for a wide variety of enzymatic activities.

Original languageEnglish (US)
Pages (from-to)10635-10642
Number of pages8
JournalAnalytical Chemistry
Volume85
Issue number22
DOIs
StatePublished - Nov 19 2013

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Enzyme activity
Mass spectrometry
Peptides
Self assembled monolayers
Gene expression
Oligonucleotides
Lysine
Labels
Cells

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

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abstract = "The development of arrays that can profile molecular activities in cells is important to understanding signaling pathways in normal and pathological settings. While oligonucleotide arrays are now routinely used to profile global gene expression, there is still a lack of tools for profiling enzyme activities in cell lysates. This paper describes the combination of peptide arrays formed on self-assembled monolayers and mass spectrometry to provide a label-free approach for identifying patterns of enzyme activities in cell lysates. The approach is demonstrated by profiling lysine deacetylase (KDAC) activities in cell lysates of the CHRF megakaryocytic (Mk) cell line. Class-specific deacetylase inhibitors were used to show that terminal Mk differentiation of CHRF cells is marked by a pronounced decrease in sirtuin activity and by little change in activity of KDACs 1-11. This work establishes a platform that can be used to identify changes in global activity profiles of cell lysates for a wide variety of enzymatic activities.",
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Profiling deacetylase activities in cell lysates with peptide arrays and SAMDI mass spectrometry. / Kuo, Hsin Yu; Deluca, Teresa A.; Miller, William M; Mrksich, Milan.

In: Analytical Chemistry, Vol. 85, No. 22, 19.11.2013, p. 10635-10642.

Research output: Contribution to journalReview article

TY - JOUR

T1 - Profiling deacetylase activities in cell lysates with peptide arrays and SAMDI mass spectrometry

AU - Kuo, Hsin Yu

AU - Deluca, Teresa A.

AU - Miller, William M

AU - Mrksich, Milan

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AB - The development of arrays that can profile molecular activities in cells is important to understanding signaling pathways in normal and pathological settings. While oligonucleotide arrays are now routinely used to profile global gene expression, there is still a lack of tools for profiling enzyme activities in cell lysates. This paper describes the combination of peptide arrays formed on self-assembled monolayers and mass spectrometry to provide a label-free approach for identifying patterns of enzyme activities in cell lysates. The approach is demonstrated by profiling lysine deacetylase (KDAC) activities in cell lysates of the CHRF megakaryocytic (Mk) cell line. Class-specific deacetylase inhibitors were used to show that terminal Mk differentiation of CHRF cells is marked by a pronounced decrease in sirtuin activity and by little change in activity of KDACs 1-11. This work establishes a platform that can be used to identify changes in global activity profiles of cell lysates for a wide variety of enzymatic activities.

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