Homologous Hevea brasiliensis REF (Hevb1) and SRPP (Hevb3) present different auto-assembling
Homologous Hevea brasiliensis REF (Hevb1) and SRPP (Hevb3) present different auto-assembling
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DOI：
10.1016/j.bbapap.2013.10.017
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发表时间：
2014-02-01
影响因子：
3.2
通讯作者：
Peruch, Frederic
中科院分区：
文献类型：
--
作者：
Berthelot, Karine;Lecomte, Sophie;Peruch, Frederic
HbREF 和 HbSRPP 是存在于橡胶颗粒上的两种巴西橡胶树蛋白，可能参与乳胶的凝固。它们的功能尚不清楚，但我们之前发现 REF 具有淀粉样蛋白特性，这在凝血过程中可能特别令人感兴趣。首先，我们确认橡胶乳胶中的同源蛋白和主要蛋白 REF 和 SAPP 不是糖蛋白。在这项工作中，我们研究了蛋白质相互作用的各个方面：聚集、自动组装、酵母和红细胞凝集、各种生化（PAGE、光谱学、显微镜）、生物物理（DLS、椭圆光度法）和结构（TEM、ATR-FTIR、PM-IRRAS）方法的相互作用。我们证明这两种蛋白质会自动组装成不同的聚集状态：REF聚合为富含β-折叠的淀粉样蛋白，并快速形成大聚集体（>μm），而SRPP在溶液中自动组装成更球状的稳定纳米多聚体。然而，两种蛋白质能够相互作用，并且 SAPP 可能抑制 REF 的淀粉样蛋白生成。 REF 还能够与酵母和红细胞的膜相互作用，导致它们凝集。此外，我们还表明，REF 和 SRPP 均不具有抗菌活性，但它们对膜的活性已得到明确证明。我们可能怀疑这些聚集特性，即使它们明显不同，也可能在凝结过程中发生，此时膜不稳定。蛋白质与膜的相互作用有助于胶乳的胶体稳定性，而蛋白质与蛋白质的相互作用则通过将橡胶颗粒聚集在一起或最终破坏颗粒单膜来促进凝固过程。 (C) 2013 Elsevier B.V. 保留所有权利。
HbREF and HbSRPP are two Hevea brasiliensis proteins present on rubber particles, and probably involved in the coagulation of latex. Their function is unclear, but we previously discovered that REF had amyloid properties, which could be of particular interest during the coagulation process. First, we confirmed that REF and SAPP, homologous and principal proteins in hevea latex, are not glycoproteins. In this work, we investigated various aspects of protein interactions: aggregation, auto-assembling, yeast and erythrocyte agglutination, co-interactions by various biochemical (PAGE, spectroscopy, microscopy), biophysical (DLS, ellipsometry) and structural (TEM, ATR-FTIR, PM-IRRAS) approaches. We demonstrated that both proteins are auto-assembling into different aggregative states: REF polymerizes as an amyloid rich in beta-sheets and forms quickly large aggregates (>mu m), whereas SRPP auto-assembles in solution into stable nanomultimers of a more globular nature. Both proteins are however able to interact together, and SAPP may inhibit the amyloidogenesis of REF. REF is also able to interact with the membranes of yeasts and erythrocytes, leading to their agglutination. In addition, we also showed that both REF and SRPP did not have antimicrobial activity, whereas their activity on membranes has been clearly evidenced. We may suspect that these aggregative properties, even though they are clearly different, may occur during coagulation, when the membrane is destabilized. The interaction of proteins with membranes could help in the colloidal stability of latex, whereas the protein-protein interactions would contribute to the coagulation process, by bringing rubber particles together or eventually disrupting the particle monomembranes. (C) 2013 Elsevier B.V. All rights reserved.