The structural basis for regulation of the glutathione transporter Ycf1 by regulatory domain phosphorylation.
The structural basis for regulation of the glutathione transporter Ycf1 by regulatory domain phosphorylation.
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DOI：
10.1038/s41467-022-28811-w
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发表时间：
2022-03-11
影响因子：
16.6
通讯作者：
Tomasiak TM
中科院分区：
文献类型：
--
作者：
Khandelwal NK;Millan CR;Zangari SI;Avila S;Williams D;Thaker TM;Tomasiak TM
酵母镉因子1（Ycf 1）将重金属和谷胱甘肽隔离到液泡中以对抗细胞应激。Ycf 1属于ATP结合盒C亚家族（ABCC）的转运蛋白，其中许多是通过磷酸化内在无序结构域来调节的。磷酸化的调节机制仍然知之甚少。在这里，我们报告了两个cryo-EM结构的Ycf 1在3.4 μ m和4.0 μ m的分辨率在面向内的开放构象，捕获以前未观察到的有序状态的内在无序的监管结构域（R-域）。R结构域磷酸化是明显的，并诱导拓扑结构，促进与核苷酸结合结构域1（NBD 1）和Lasso基序的静电和疏水相互作用。这些相互作用在结构之间保持恒定，并且与NBD 1/R结构域复合物的刚体运动相关。生化数据进一步表明，R-结构域磷酸化重组Ycf 1架构，并需要最大的ATP酶活性。总之，我们提供了R-结构域如何控制ABCC转运蛋白活性的见解。Ycf 1是C家族成员ATP结合盒（ABC）转运蛋白，在酵母中转运谷胱甘肽和谷胱甘肽-金属复合物。在这里，作者使用冷冻EM和生化分析来显示内在无序的调节结构域（R结构域）如何通过与核苷酸结合结构域接合来控制磷酸化后的活性。
Yeast Cadmium Factor 1 (Ycf1) sequesters heavy metals and glutathione into the vacuole to counter cell stress. Ycf1 belongs to the ATP binding cassette C-subfamily (ABCC) of transporters, many of which are regulated by phosphorylation on intrinsically-disordered domains. The regulatory mechanism of phosphorylation is still poorly understood. Here, we report two cryo-EM structures of Ycf1 at 3.4 Å and 4.0 Å resolution in inward-facing open conformations that capture previously unobserved ordered states of the intrinsically disordered regulatory domain (R-domain). R-domain phosphorylation is clearly evident and induces a topology promoting electrostatic and hydrophobic interactions with Nucleotide Binding Domain 1 (NBD1) and the Lasso motif. These interactions stay constant between the structures and are related by rigid body movements of the NBD1/R-domain complex. Biochemical data further show R-domain phosphorylation reorganizes the Ycf1 architecture and is required for maximal ATPase activity. Together, we provide insights into how R-domains control ABCC transporter activity. Ycf1, a C-family member ATP Binding Cassette (ABC) transporter, transports glutathione and glutathione-metal complexes in yeast. Here the authors use cryo-EM and biochemical analysis to show how an intrinsically-disordered regulatory domain (R-domain) controls activity upon phosphorylation by engaging with a Nucleotide Binding Domain.
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影响因子：
48
作者：
Kucukelbir, Alp;Sigworth, Fred J.;Tagare, Hemant D.
通讯作者：
Tagare, Hemant D.
影响因子：
3.5
作者：
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影响因子：
64.8
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Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
16.8
作者：
Baker, Jennifer M. R.;Hudson, Rhea P.;Forman-Kay, Julie D.
通讯作者：
Forman-Kay, Julie D.