Identification of an allosteric signaling network within Tec family kinases.
Identification of an allosteric signaling network within Tec family kinases.
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DOI：
10.1016/j.jmb.2010.08.035
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发表时间：
2010-10-22
影响因子：
5.6
通讯作者：
Andreotti AH
中科院分区：
文献类型：
--
作者：
Joseph RE;Xie Q;Andreotti AH
Tec家族激酶是主要在造血细胞中起作用的酪氨酸激酶。Tec激酶的催化活性受到激酶结构域外的调节结构域的积极影响。目前缺乏全长Tec激酶结构，使我们无法理解这些正调控信号如何传递到激酶结构域。最近，激酶内的保守结构，“调节脊柱”，已被确定为组装和拆卸的激酶之间的开关，其活性和非活性状态。在这里，我们定义的残基，包括在Tec激酶的调节脊柱。与先前表征的系统相比，Tec激酶含有延伸的调控脊柱，其包括Tec激酶的C-螺旋内的保守甲硫氨酸和SH 2-激酶接头内的保守色氨酸。这种延伸的调节刺形成了一个管道，用于将Tec激酶的调节结构域的存在传递到催化结构域。我们进一步表明，突变的看门人残基在边缘的监管脊柱稳定的监管脊柱，导致组成型活性激酶结构域。重要的是，调控脊柱在这个看门突变体中预先组装，使得激活环上的磷酸化对其活性是不必要的。此外，我们表明，破坏Btk R544的激活环和Btk E445的C-螺旋之间的保守的静电相互作用也有助于组装的监管脊柱。因此，延伸的调节棘是维持Tec激酶活性的关键结构。
The Tec family kinases are tyrosine kinases that function primarily in hematopoietic cells. The catalytic activity of the Tec kinases is positively influenced by the regulatory domains outside of the kinase domain. The current lack of a full-length Tec kinase structure leaves a void in our understanding of how these positive regulatory signals are transmitted to the kinase domain. Recently, a conserved structure within kinases, the ‘regulatory spine’, has been identified that assembles and disassembles as a kinase switches between its active and inactive states. Here we define the residues that comprise the regulatory spine within Tec kinases. Compared to previously characterized systems, the Tec kinases contain an extended regulatory spine that includes a conserved methionine within the C-helix and a conserved tryptophan within the SH2-kinase linker of Tec kinases. This extended regulatory spine forms a conduit for transmitting the presence of the regulatory domains of Tec kinases to the catalytic domain. We further show that mutation of the gatekeeper residue at the edge of the regulatory spine stabilizes the regulatory spine resulting in a constitutively active kinase domain. Importantly, the regulatory spine is preassembled in this gatekeeper mutant rendering phosphorylation on the activation loop unnecessary for its activity. Moreover, we show that the disruption of the conserved electrostatic interaction between Btk R544 on the activation loop and Btk E445 on the C-helix also aids in the assembly of the regulatory spine. Thus, the extended regulatory spine is a key structure that is critical for maintaining the activity of Tec kinases.
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影响因子：
8.7
作者：
Joseph RE;Andreotti AH
通讯作者：
Andreotti AH
影响因子：
2.9
作者：
Joseph, Raji E.;Min, Lie;Andreotti, Amy H.
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作者：
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通讯作者：
Hubbard, SR
影响因子：
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作者：
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通讯作者：
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