Structural basis for allosteric regulation of human phosphofructokinase-1.
Structural basis for allosteric regulation of human phosphofructokinase-1.
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人磷酸果糖激酶 1 变构调节的结构基础。
DOI：
10.1101/2024.03.15.585110
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Webb,BradleyA
中科院分区：
文献类型：
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作者：
Lynch,EricM;Hansen,Heather;Salay,Lauren;Cooper,Madison;Timr,Stepan;Kollman,JustinM;Webb,BradleyA
磷酸果糖激酶-1 （PFK1）催化糖酵解的限速步骤，将葡萄糖转化为细胞能量。PFK1受到高度调控，以响应细胞不断变化的能量需求。在细菌中，PFK1调控的结构基础是变构学的教科书范例；低能量和高能量的分子信号分别促进活性r态和非活性t态构象之间的转变。然而，关于真核生物PFK1调控的结构基础知之甚少。在这里，我们通过低温电镜测定了人类肝脏PFK1 （PFKL）在R和t状态下的结构，为真核PFK1变构调节机制提供了见解。t态结构揭示了细菌和真核酶之间的构象差异，ATP在多个位点结合的变构抑制机制，以及c端在稳定t态中的自抑制作用。我们还确定了确定高阶组装机制的PFKL丝的结构，并证明这些结构是细胞中PFKL高阶组装所必需的。
Phosphofructokinase-1 (PFK1) catalyzes the rate-limiting step of glycolysis, committing glucose to conversion into cellular energy. PFK1 is highly regulated to respond to the changing energy needs of the cell. In bacteria, the structural basis of PFK1 regulation is a textbook example of allostery; molecular signals of low and high cellular energy promote transition between an active R-state and inactive T-state conformation, respectively. Little is known, however, about the structural basis for regulation of eukaryotic PFK1. Here, we determine structures of the human liver isoform of PFK1 (PFKL) in the R- and T-state by cryoEM, providing insight into eukaryotic PFK1 allosteric regulatory mechanisms. The T-state structure reveals conformational differences between the bacterial and eukaryotic enzyme, the mechanisms of allosteric inhibition by ATP binding at multiple sites, and an autoinhibitory role of the C-terminus in stabilizing the T-state. We also determine structures of PFKL filaments that define the mechanism of higher-order assembly and demonstrate that these structures are necessary for higher-order assembly of PFKL in cells.