A p38α-selective chemosensor for use in unfractionated cell lysates

Clifford I Stains, Elvedin Luković, Barbara Imperiali

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Recent efforts have identified the p38α Ser/Thr kinase as a potential target for the treatment of inflammatory diseases as well as non-small cell lung carcinoma. Despite the significance of p38α, no direct activity probe compatible with cell lysate analysis exists. Instead, proxies for kinase activation, such as phosphospecific antibodies, which do not distinguish between p38 isoforms, are often used. Our laboratory has recently developed a sulfonamido-oxine (Sox) fluorophore that undergoes a significant increase in fluorescence in response to phosphorylation at a proximal residue, allowing for real-time activity measurements. Herein we report the rational design of a p38α-selective chemosensor using this approach. We have validated the selectivity of this sensor using specific inhibitors and immunodepletions and show that p38α activity can be monitored in crude lysates from a variety of cell lines, allowing for the potential use of this sensor in both clinical and basic science research applications.

Original languageEnglish (US)
Pages (from-to)101-105
Number of pages5
JournalACS Chemical Biology
Volume6
Issue number1
DOIs
StatePublished - Jan 21 2011

Fingerprint

Phosphotransferases
Cells
Phospho-Specific Antibodies
Oxyquinoline
Phosphorylation
Fluorophores
Sensors
Proxy
Non-Small Cell Lung Carcinoma
Protein Isoforms
Fluorescence
Chemical activation
Cell Line
Research
Therapeutics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

A p38α-selective chemosensor for use in unfractionated cell lysates. / Stains, Clifford I; Luković, Elvedin; Imperiali, Barbara.

In: ACS Chemical Biology, Vol. 6, No. 1, 21.01.2011, p. 101-105.

Research output: Contribution to journalArticle

Stains, Clifford I ; Luković, Elvedin ; Imperiali, Barbara. / A p38α-selective chemosensor for use in unfractionated cell lysates. In: ACS Chemical Biology. 2011 ; Vol. 6, No. 1. pp. 101-105.
@article{22571e3f666a45728de02ed6d3ac602b,
title = "A p38α-selective chemosensor for use in unfractionated cell lysates",
abstract = "Recent efforts have identified the p38α Ser/Thr kinase as a potential target for the treatment of inflammatory diseases as well as non-small cell lung carcinoma. Despite the significance of p38α, no direct activity probe compatible with cell lysate analysis exists. Instead, proxies for kinase activation, such as phosphospecific antibodies, which do not distinguish between p38 isoforms, are often used. Our laboratory has recently developed a sulfonamido-oxine (Sox) fluorophore that undergoes a significant increase in fluorescence in response to phosphorylation at a proximal residue, allowing for real-time activity measurements. Herein we report the rational design of a p38α-selective chemosensor using this approach. We have validated the selectivity of this sensor using specific inhibitors and immunodepletions and show that p38α activity can be monitored in crude lysates from a variety of cell lines, allowing for the potential use of this sensor in both clinical and basic science research applications.",
author = "Stains, {Clifford I} and Elvedin Luković and Barbara Imperiali",
year = "2011",
month = "1",
day = "21",
doi = "10.1021/cb100230y",
language = "English (US)",
volume = "6",
pages = "101--105",
journal = "ACS Chemical Biology",
issn = "1554-8929",
publisher = "American Chemical Society",
number = "1",

}

TY - JOUR

T1 - A p38α-selective chemosensor for use in unfractionated cell lysates

AU - Stains, Clifford I

AU - Luković, Elvedin

AU - Imperiali, Barbara

PY - 2011/1/21

Y1 - 2011/1/21

N2 - Recent efforts have identified the p38α Ser/Thr kinase as a potential target for the treatment of inflammatory diseases as well as non-small cell lung carcinoma. Despite the significance of p38α, no direct activity probe compatible with cell lysate analysis exists. Instead, proxies for kinase activation, such as phosphospecific antibodies, which do not distinguish between p38 isoforms, are often used. Our laboratory has recently developed a sulfonamido-oxine (Sox) fluorophore that undergoes a significant increase in fluorescence in response to phosphorylation at a proximal residue, allowing for real-time activity measurements. Herein we report the rational design of a p38α-selective chemosensor using this approach. We have validated the selectivity of this sensor using specific inhibitors and immunodepletions and show that p38α activity can be monitored in crude lysates from a variety of cell lines, allowing for the potential use of this sensor in both clinical and basic science research applications.

AB - Recent efforts have identified the p38α Ser/Thr kinase as a potential target for the treatment of inflammatory diseases as well as non-small cell lung carcinoma. Despite the significance of p38α, no direct activity probe compatible with cell lysate analysis exists. Instead, proxies for kinase activation, such as phosphospecific antibodies, which do not distinguish between p38 isoforms, are often used. Our laboratory has recently developed a sulfonamido-oxine (Sox) fluorophore that undergoes a significant increase in fluorescence in response to phosphorylation at a proximal residue, allowing for real-time activity measurements. Herein we report the rational design of a p38α-selective chemosensor using this approach. We have validated the selectivity of this sensor using specific inhibitors and immunodepletions and show that p38α activity can be monitored in crude lysates from a variety of cell lines, allowing for the potential use of this sensor in both clinical and basic science research applications.

UR - http://www.scopus.com/inward/record.url?scp=79251482450&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79251482450&partnerID=8YFLogxK

U2 - 10.1021/cb100230y

DO - 10.1021/cb100230y

M3 - Article

VL - 6

SP - 101

EP - 105

JO - ACS Chemical Biology

JF - ACS Chemical Biology

SN - 1554-8929

IS - 1

ER -