Zilkha Neurogenetic Institute, University of Southern California, Los Angeles, California 90033, USA.
J Biol Chem. 2012 Feb 17;287(8):5235-41. doi: 10.1074/jbc.M111.327817. Epub 2011 Dec 20.
Misfolding and amyloid fibril formation by human islet amyloid polypeptide (hIAPP) are thought to be important in the pathogenesis of type 2 diabetes, but the structures of the misfolded forms remain poorly understood. Here we developed an approach that combines site-directed spin labeling with continuous wave and pulsed EPR to investigate local secondary structure and to determine the relative orientation of the secondary structure elements with respect to each other. These data indicated that individual hIAPP molecules take up a hairpin fold within the fibril. This fold contains two β-strands that are much farther apart than expected from previous models. Atomistic structural models were obtained using computational refinement with EPR data as constraints. The resulting family of structures exhibited a left-handed helical twist, in agreement with the twisted morphology observed by electron microscopy. The fibril protofilaments contain stacked hIAPP monomers that form opposing β-sheets that twist around each other. The two β-strands of the monomer adopt out-of-plane positions and are staggered by about three peptide layers (∼15 Å). These results provide a mechanism for hIAPP fibril formation and could explain the remarkable stability of the fibrils. Thus, the structural model serves as a starting point for understanding and preventing hIAPP misfolding.
人胰岛淀粉样多肽(hIAPP)的错误折叠和淀粉样纤维形成被认为在 2 型糖尿病的发病机制中很重要,但错误折叠形式的结构仍知之甚少。在这里,我们开发了一种结合定点自旋标记与连续波和脉冲 EPR 的方法来研究局部二级结构,并确定二级结构元件之间的相对取向。这些数据表明,单个 hIAPP 分子在纤维中采取发夹折叠。这种折叠包含两个 β-链,它们之间的距离比以前的模型预期的要远得多。使用 EPR 数据作为约束的计算精修获得原子结构模型。得到的结构家族表现出左手螺旋扭曲,与电子显微镜观察到的扭曲形态一致。纤维原丝包含堆叠的 hIAPP 单体,形成相互扭曲的相反 β-片层。单体的两条 β-链采用离域位置,交错约三个肽层(约 15 Å)。这些结果为 hIAPP 纤维形成提供了一种机制,并可以解释纤维的惊人稳定性。因此,该结构模型为理解和预防 hIAPP 错误折叠提供了一个起点。