An optogenetic tool to recruit individual PKC isozymes to the cell surface and promote specific phosphorylation of membrane proteins.
An optogenetic tool to recruit individual PKC isozymes to the cell surface and promote specific phosphorylation of membrane proteins.
复制标题
DOI：
10.1016/j.jbc.2022.101893
复制
发表时间：
2022-05
影响因子：
4.8
通讯作者：
Logothetis, Diomedes E.
中科院分区：
文献类型：
--
作者：
Gada, Kirin D.;Kawano, Takeharu;Plant, Leigh D.;Logothetis, Diomedes E.
PKC家族由几种密切相关的激酶组成。这些酶通过丝氨酸和/或苏氨酸上羟基的磷酸化来调节蛋白质的功能。单个PKC同工酶的选择性激活已被证明具有挑战性，因为缺乏特异性激活剂分子。在这里，我们开发了一种光遗传学蓝光激活的PKC同工酶，利用光敏隐花色素-2（CYP 12）和转录因子钙和整合素结合蛋白1（CIB 1）的N末端（CYP 12结合结构域的N末端区域）之间的基于植物的二聚化系统。我们发现，标记PKC同工酶的催化结构域的p53 2可以有效地促进其易位到细胞表面后，蓝光照射。我们使用PKCε证明了该系统，并表明这导致K+通道（G蛋白门控内向整流K+通道1和4）的强烈激活，先前显示由PKCε激活。我们预计，这种方法可以用于其他PKC亚型，提供一个可靠的和直接的刺激靶向膜蛋白磷酸化的相关PKC。
The PKC family consists of several closely related kinases. These enzymes regulate the function of proteins through the phosphorylation of hydroxyl groups on serines and/or threonines. The selective activation of individual PKC isozymes has proven challenging because of a lack of specific activator molecules. Here, we developed an optogenetic blue light–activated PKC isozyme that harnesses a plant-based dimerization system between the photosensitive cryptochrome-2 (CRY2) and the N terminus of the transcription factor calcium and integrin-binding protein 1 (CIB1) (N-terminal region of the CRY2-binding domain of CIB1). We show that tagging CRY2 with the catalytic domain of PKC isozymes can efficiently promote its translocation to the cell surface upon blue light exposure. We demonstrate this system using PKCε and show that this leads to robust activation of a K+ channel (G protein–gated inwardly rectifying K+ channels 1 and 4), previously shown to be activated by PKCε. We anticipate that this approach can be utilized for other PKC isoforms to provide a reliable and direct stimulus for targeted membrane protein phosphorylation by the relevant PKCs.
登录
查看更多内容
影响因子：
56.9
作者：
Liu, Hongtao;Yu, Xuhong;Lin, Chentao
通讯作者：
Lin, Chentao
DOI：
10.1073/pnas.1211305109
发表时间：
2012-08-28
影响因子：
11.1
作者：
Idevall-Hagren, Olof;Dickson, Eamonn J.;De Camilli, Pietro
通讯作者：
De Camilli, Pietro
影响因子：
15
作者：
Schleifenbaum, A;Stier, G;Schultz, C
通讯作者：
Schultz, C
影响因子：
6.5
作者：
Kanno, Takeshi;Yamamoto, Hideyuki;Nishizaki, Tomoyuki
通讯作者：
Nishizaki, Tomoyuki
影响因子：
14.5
作者：
Newton AC
通讯作者：
Newton AC