Thiol-ene-Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins

Kaelyn E. Wilke; Erin E Carlson

doi:10.1007/978-1-4939-3049-4_1

Thiol-ene-Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins

Kaelyn E. Wilke, Erin E Carlson

Chemistry (Twin Cities)

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Abstract

The adenosine triphosphate (ATP) analogue adenosine 5'-O-(3-thiotriphosphate) (ATPγS) has been applied as a tool to study kinase-substrate phosphorylation. Not only does the transfer of a thiophosphate group represent a unique modification amid the phosphoproteome, but it can also be stable to phosphatase activity. However, detection of this species is complicated due to the similar chemical reactivity of thiophosphate and proteinaceous thiols. Here, we describe a novel method for detection of protein thiophosphorylation utilizing the thiol-ene reaction. By first chemoselectively capping protein thiols through radical chemistry, kinase-catalyzed thiophosphorylation can be visualized specifically.

Original language	English (US)
Pages (from-to)	3-15
Number of pages	13
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	1355
DOIs	https://doi.org/10.1007/978-1-4939-3049-4_1
State	Published - Jan 1 2016

Keywords

ATPγS
Kinase
Phosphoprotein
Substrate
Thiol capping
Thiol-ene
Thiophosphorylation

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title = "Thiol-ene-Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins",

abstract = "The adenosine triphosphate (ATP) analogue adenosine 5'-O-(3-thiotriphosphate) (ATPγS) has been applied as a tool to study kinase-substrate phosphorylation. Not only does the transfer of a thiophosphate group represent a unique modification amid the phosphoproteome, but it can also be stable to phosphatase activity. However, detection of this species is complicated due to the similar chemical reactivity of thiophosphate and proteinaceous thiols. Here, we describe a novel method for detection of protein thiophosphorylation utilizing the thiol-ene reaction. By first chemoselectively capping protein thiols through radical chemistry, kinase-catalyzed thiophosphorylation can be visualized specifically. ",

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T1 - Thiol-ene-Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins

AU - Wilke, Kaelyn E.

AU - Carlson, Erin E

PY - 2016/1/1

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N2 - The adenosine triphosphate (ATP) analogue adenosine 5'-O-(3-thiotriphosphate) (ATPγS) has been applied as a tool to study kinase-substrate phosphorylation. Not only does the transfer of a thiophosphate group represent a unique modification amid the phosphoproteome, but it can also be stable to phosphatase activity. However, detection of this species is complicated due to the similar chemical reactivity of thiophosphate and proteinaceous thiols. Here, we describe a novel method for detection of protein thiophosphorylation utilizing the thiol-ene reaction. By first chemoselectively capping protein thiols through radical chemistry, kinase-catalyzed thiophosphorylation can be visualized specifically.

AB - The adenosine triphosphate (ATP) analogue adenosine 5'-O-(3-thiotriphosphate) (ATPγS) has been applied as a tool to study kinase-substrate phosphorylation. Not only does the transfer of a thiophosphate group represent a unique modification amid the phosphoproteome, but it can also be stable to phosphatase activity. However, detection of this species is complicated due to the similar chemical reactivity of thiophosphate and proteinaceous thiols. Here, we describe a novel method for detection of protein thiophosphorylation utilizing the thiol-ene reaction. By first chemoselectively capping protein thiols through radical chemistry, kinase-catalyzed thiophosphorylation can be visualized specifically.

KW - ATPγS

KW - Kinase

KW - Phosphoprotein

KW - Substrate

KW - Thiol capping

KW - Thiol-ene

KW - Thiophosphorylation

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U2 - 10.1007/978-1-4939-3049-4_1

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JO - Methods in molecular biology (Clifton, N.J.)

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ER -

Thiol-ene-Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins

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