Guijarro J I, Sunde M, Jones J A, Campbell I D, Dobson C M
Oxford Centre for Molecular Sciences, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom.
Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4224-8. doi: 10.1073/pnas.95.8.4224.
The SH3 domain is a well characterized small protein module with a simple fold found in many proteins. At acid pH, the SH3 domain (PI3-SH3) of the p85alpha subunit of bovine phosphatidylinositol 3-kinase slowly forms a gel that consists of typical amyloid fibrils as assessed by electron microscopy, a Congo red binding assay, and x-ray fiber diffraction. The soluble form of PI3-SH3 at acid pH (the A state by a variety of techniques) from which fibrils are generated has been characterized. Circular dichroism in the far- and near-UV regions and 1H NMR indicate that the A state is substantially unfolded relative to the native protein at neutral pH. NMR diffusion measurements indicate, however, that the effective hydrodynamic radius of the A state is only 23% higher than that of the native protein and is 20% lower than that of the protein denatured in 3.5 M guanidinium chloride. In addition, the A state binds the hydrophobic dye 1-anilinonaphthalene-8-sulfonic acid, which suggests that SH3 in this state has a partially formed hydrophobic core. These results indicate that the A state is partially folded and support the hypothesis that partially folded states formed in solution are precursors of amyloid deposition. Moreover, that this domain aggregates into amyloid fibrils suggests that the potential for amyloid deposition may be a common property of proteins, and not only of a few proteins associated with disease.
SH3结构域是一种特征明确的小蛋白模块,具有简单的折叠结构,存在于许多蛋白质中。在酸性pH条件下,牛磷脂酰肌醇3激酶p85α亚基的SH3结构域(PI3-SH3)会缓慢形成一种凝胶,通过电子显微镜、刚果红结合试验和X射线纤维衍射评估,该凝胶由典型的淀粉样纤维组成。已对酸性pH条件下PI3-SH3的可溶形式(通过多种技术确定为A状态)进行了表征,淀粉样纤维即由此状态产生。远紫外和近紫外区域的圆二色性以及1H NMR表明,相对于中性pH条件下的天然蛋白质,A状态基本处于未折叠状态。然而,NMR扩散测量表明,A状态的有效流体动力学半径仅比天然蛋白质高23%,比在3.5 M氯化胍中变性的蛋白质低20%。此外,A状态能结合疏水性染料1-苯胺基萘-8-磺酸,这表明该状态下的SH3具有部分形成的疏水核心。这些结果表明A状态是部分折叠的,并支持溶液中形成的部分折叠状态是淀粉样沉积前体的假说。此外,该结构域聚集成淀粉样纤维表明,淀粉样沉积的可能性可能是蛋白质的普遍特性,而不仅是与疾病相关的少数蛋白质的特性。