A semisynthetic epitope for kinase substrates.

A semisynthetic epitope for kinase substrates. Research Abstract Details 

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A semisynthetic epitope for kinase substrates. Abstract Text:

Jasmina J Allen,Manqing Li,Craig S Brinkworth,Jennifer L Paulson,Dan Wang,Anette ,Wen-Hai Chou,Roger J Davis,Alma L Burlingame,Robert O Messing,Carol D Katayama,Stephen M Hedrick,Kevan M Shokat,

The ubiquitous nature of protein phosphorylation makes it challenging to map kinase-substrate relationships, which is a necessary step toward defining signaling network architecture. To trace the activity of individual kinases, we developed a semisynthetic reaction scheme, which results in the affinity tagging of substrates of the kinase in question. First, a kinase, engineered to use a bio-orthogonal ATPgammaS analog, catalyzes thiophosphorylation of its direct substrates. Second, alkylation of thiophosphorylated serine, threonine or tyrosine residues creates an epitope for thiophosphate ester-specific antibodies. We demonstrated the generality of semisynthetic epitope construction with 13 diverse kinases: JNK1, p38alpha MAPK, Erk1, Erk2, Akt1, PKCdelta, PKCepsilon, Cdk1/cyclinB, CK1, Cdc5, GSK3beta, Src and Abl. Application of this approach, in cells isolated from a mouse that expressed endogenous levels of an analog-specific (AS) kinase (Erk2), allowed purification of a direct Erk2 substrate.

A semisynthetic epitope for kinase substrates. Publishing Authors By Initials

JJ Allen,M Li,CS Brinkworth,JL Paulson,D Wang,A ,WH Chou,RJ Davis,AL Burlingame,RO Messing,CD Katayama,SM Hedrick,KM Shokat,

For similar organic chemicals: organophosphorus compounds: organothiophosphorus compounds: thiophosphoric acid esters research abstracts see: organic chemicals: organophosphorus compounds: organothiophosphorus compounds: thiophosphoric acid esters research

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A semisynthetic epitope for kinase substrates. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Nature methods

VOLUME: 4

Page Numbers: 511-6

Journal Abbreviation: Nat. Methods

ISSN: 1548-7091

DAY: 7

MONTH: 05

YEAR: 2007

A semisynthetic epitope for kinase substrates. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 101215604

A semisynthetic epitope for kinase substrates. Keywords Mesh Terms:

KEYWORDS: Thiophosphoric Acid Esters

MESH TERMS: metabolism

Chemical & Substance for Abstract: A semisynthetic epitope for kinase substrates. Information

Substance Name: Extracellular Signal-Regulated MAP Kinas

Registry Number: EC 2.7.1.37

Grant and Affiliation Information for A semisynthetic epitope for kinase substrates.

AFFILIATION: Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, San Francisco, California 94143, USA.

Country: United States

AGENCY: United States NCRR

GRANT: RR015804

ACRONYM: RR

MEDLINETA: Nat Methods

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