Synthesis And Application Of Atp Analogs For Phosphorylation-Dependent Kinase-Substrate Crosslinking
type: CreativeWork
description: Phosphorylation is an important post-translational modification that plays a key role in a variety of signaling cascades and cellular functions. Kinases phosphorylate protein substrates in a highly regulated manner and are promiscuous. Understanding kinase-substrate specificity has been challenging and there is a need for new chemical tools. To this end we developed -phosphate modified ATP photocrosslinking analogs ATP-ArN3 and ATP-BP, that crosslink substrate and kinase in a phosphorylation dependent manner. We have successfully demonstrated that ATP-ArN3 and ATP-BP can be used with natural kinase and substrates using cell lysates in vitro. We used our approach to identify novel kinases of p53. One powerful feature of this methodology is we can obtain an atomic level snapshot of the interactions between proteins, when coupled with analytical techniques like Mass Spectrometry (MS). These tools will help us in validating our understanding of protein-protein interactions, their role in signaling pathways and functioning of the cell.
abstract: Phosphorylation is an important post-translational modification that plays a key role in a variety of signaling cascades and cellular functions. Kinases phosphorylate protein substrates in a highly regulated manner and are promiscuous. Understanding kinase-substrate specificity has been challenging and there is a need for new chemical tools. To this end we developed -phosphate modified ATP photocrosslinking analogs ATP-ArN3 and ATP-BP, that crosslink substrate and kinase in a phosphorylation dependent manner. We have successfully demonstrated that ATP-ArN3 and ATP-BP can be used with natural kinase and substrates using cell lysates in vitro. We used our approach to identify novel kinases of p53. One powerful feature of this methodology is we can obtain an atomic level snapshot of the interactions between proteins, when coupled with analytical techniques like Mass Spectrometry (MS). These tools will help us in validating our understanding of protein-protein interactions, their role in signaling pathways and functioning of the cell.
author: Garre Venkata Raghavendra, Satish Kumar
keywords: ATP, crosslinking, kinases, p53, phosphorylation, post-translation modifications, Analytical Chemistry, Biochemistry, Organic Chemistry
date published: 2013-01-01T08:00:00Z
date modified: 2026-07-24T05:59:33Z
provider: Wayne State University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Wayne State University