Self-Assembly of Human Profilin-1 Detected by Carr-Purcell-Meiboom-Gill Nuclear Magnetic Resonance (CPMG NMR) Spectroscopy
Self-Assembly of Human Profilin-1 Detected by Carr-Purcell-Meiboom-Gill Nuclear Magnetic Resonance (CPMG NMR) Spectroscopy
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DOI：
10.1021/acs.biochem.6b01263
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发表时间：
2017-02-07
期刊：
影响因子：
2.9
通讯作者：
Kay, Lewis E.
中科院分区：
文献类型：
--
作者：
Rennella, Enrico;Sekhar, Ashok;Kay, Lewis E.
细胞中的蛋白质寡聚化对健康和疾病都有重要意义，因此，对蛋白质自缔合机制的理解具有重要意义。寡聚化过程的初始阶段可能难以检测，因为它们通常涉及难以通过标准生物物理方法表征的稀疏分布和瞬态的形成。使用弛豫色散核磁共振波谱，我们研究了人profilin-1，一种调节肌动蛋白聚合的蛋白质的寡聚化。我们表明，在溶液中，并在毫摩尔浓度profilin-1主要是单体。然而，浓度依赖性弛豫数据的拟合与通过多步过程生成的高阶低聚物的形成是一致的。连同晶体学数据的profilin-2，同源的蛋白质研究在这里，我们的研究结果表明，profilin-1形成一个稀疏的四聚体构象在溶液中。
Protein oligomerization in the cell has important implications for both health and disease, and an understanding of the mechanisms by which proteins can self-associate is, therefore, of critical interest. Initial stages of the oligomerization process can be hard to detect, as they often involve the formation of sparsely populated and transient states that are difficult to characterize by standard biophysical approaches. Using relaxation dispersion nuclear magnetic resonance spectroscopy, we study the oligomerization of human profilin-1, a protein that regulates the polymerization of actin. We show that in solution and at millimolar concentrations profilin-1 is predominantly monomeric. However, fits of concentration-dependent relaxation data are consistent with the formation of a higher-order oligomer that is generated via a multistep process. Together with crystallographic data for profilin-2, a homologue of the protein studied here, our results suggest that profilin-1 forms a sparsely populated tetrameric conformer in solution.