Hamelberg Donald, Shen Tongye, McCammon J Andrew
Howard Hughes Medical Institute, Center for Theoretical Biological Physics, Department of Chemistry and Biochemistry, Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093-0365, USA.
Proc Natl Acad Sci U S A. 2007 Sep 18;104(38):14947-51. doi: 10.1073/pnas.0703151104. Epub 2007 Sep 6.
Phosphorylation of proteins by kinases is the most commonly studied class of posttranslational modification, yet its structural consequences are not well understood. The human SR (serine-arginine) protein ASF/SF2 relies on the processive phosphorylation of the serine residues of eight consecutive arginine-serine (RS) dipeptide repeats at the C terminus by SRPK1 before it can be transported into the nucleus. This SR protein plays critical roles in spliceosome assembly, pre-mRNA splicing, and mRNA export, and the phosphorylation process of the RS repeats has been extensively studied experimentally. However, knowledge of the conformational changes associated with the phosphorylation of this simple sequence and how it triggers the importation of the SR protein is lacking. Here, we have carried out extensive molecular dynamics simulations to show that phosphorylation of the eight RS repeats significantly alters the peptide's conformation and leads to the formation of very stable structures that are likely to be involved in the recognition, binding, and transport of the SR protein. Specifically, we found an unusual symmetry-broken phase of conformations of the repetitive and quasi-symmetric phosphorylated peptide sequence. One of the main characteristics of these conformations is the exposed phosphate groups on the periphery, which possibly could serve as the recognition platform for the transport protein transportin-SR2.
激酶对蛋白质的磷酸化是最常被研究的一类翻译后修饰,但其结构后果尚未得到充分理解。人类SR(丝氨酸-精氨酸)蛋白ASF/SF2在被转运到细胞核之前,依赖于SRPK1对其C末端八个连续的精氨酸-丝氨酸(RS)二肽重复序列的丝氨酸残基进行持续性磷酸化。这种SR蛋白在剪接体组装、前体mRNA剪接和mRNA输出中发挥关键作用,并且RS重复序列的磷酸化过程已经得到了广泛的实验研究。然而,对于与这个简单序列的磷酸化相关的构象变化以及它如何触发SR蛋白的导入,目前还缺乏了解。在这里,我们进行了广泛的分子动力学模拟,以表明八个RS重复序列的磷酸化显著改变了肽的构象,并导致形成非常稳定的结构,这些结构可能参与SR蛋白的识别、结合和转运。具体来说,我们发现了重复且准对称的磷酸化肽序列构象的一个不寻常的对称破缺阶段。这些构象的一个主要特征是外围暴露的磷酸基团,它们可能作为转运蛋白transportin-SR2的识别平台。