An Affinity-Based, Cysteine-Specific ATP Analog for Kinase-Catalyzed Crosslinking.
An Affinity-Based, Cysteine-Specific ATP Analog for Kinase-Catalyzed Crosslinking.
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DOI：
10.1002/anie.202014047
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发表时间：
2021-04-26
期刊：
影响因子：
--
通讯作者：
Pflum MKH
中科院分区：
文献类型：
--
作者：
Fouda AE;Gamage AK;Pflum MKH
Kinases mediate cell signaling pathways by catalyzing protein phosphorylation. Irregularities in kinase activity are directly associated with disease conditions. Therefore, methods to identify substrates of a particular kinase are needed to understand signaling cascades in normal and diseased states. Photocrosslinking ATP analogs provide powerful tools to study kinases by covalently linking kinases with substrates. However, the involvement of UV light and nonspecific reactivity of current ATP-photocrosslinkers challenge kinase-substrate identification. We report here an affinity-based crosslinking ATP analog, ATP-methylacrylamide (ATP-MAc), that contains a cysteine-reactive acrylamide crosslinking group, which avoids the UV irradiation and non-specific reactivity of prior analogs. Using in vitro kinase assays, ATP-MAc acts as a kinase cosubstrate and covalently crosslinks only kinases containing cysteines in the active site. ATP-MAc was also able to crosslink cellular proteins in lysates, documenting compatibility with cell-based studies. Phosphorylation-dependent affinity-based crosslinking with ATP-MAc will have applications in kinase substrate identification to understand kinase-mediated cell signaling. Characterization of kinase-substrate pairs is an enabling strategy to understand kinase-related diseases and identify new drug targets. An affinity-based kinase-dependent crosslinker, ATP-MAc, is introduced to capture kinase-substrate interactions under physiological conditions. Unlike prior ATP-photocrosslinking analogs, ATP-MAc covalently links only cysteine-containing kinases with their substrates without the need for UV light.
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影响因子：
16.6
作者：
Suwal, Sujit;Pflum, Mary Kay H.
通讯作者：
Pflum, Mary Kay H.
影响因子：
2
作者：
Murale DP;Hong SC;Haque MM;Lee JS
通讯作者：
Lee JS
DOI：
10.1016/s0960-0760(01)00190-x
发表时间：
2002-02-01
影响因子：
4.1
作者：
Filardo, EJ
通讯作者：
Filardo, EJ
影响因子：
15
作者：
Maly, DJ;Allen, JA;Shokat, KM
通讯作者：
Shokat, KM
影响因子：
16
作者：
Runkle KB;Kharbanda A;Stypulkowski E;Cao XJ;Wang W;Garcia BA;Witze ES
通讯作者：
Witze ES