Stability and shape of hepatitis B virus capsids in vacuo.
Stability and shape of hepatitis B virus capsids in vacuo.
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DOI：
10.1002/anie.200802410
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发表时间：
2008-08
影响因子：
--
通讯作者：
C. Uetrecht;C. Versluis;N. Watts;P. Wingfield;A. Steven;A. Heck
中科院分区：
文献类型：
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作者：
C. Uetrecht;C. Versluis;N. Watts;P. Wingfield;A. Steven;A. Heck
B型肝炎病毒（HBV）是人类肝脏疾病的主要原因[1]，其非感染性衣壳对于纳米技术（包括药物递送应用）是有意义的。这些颗粒的精确生物物理表征不仅对这些应用非常重要，而且还因为它可以进一步了解病毒的复制周期和组装途径，从而有助于未来药物的开发。[2，3] HBV衣壳蛋白（cp）在体内和体外形成两种大小的二十面体衣壳（其中三角测量数T = 3和T = 4[4]包含180和240个子单元，衣壳蛋白具有两个结构域--核心结构域[9]在这些结构域中，核心结构域对于衣壳的组装是必需的，也是足够的。接头的长度和组装发生的条件决定了获得的T = 3和T = 4衣壳的比例。[10]衣壳蛋白二聚体通过分子间四螺旋束[11-13]和束内的二硫键（Cys 61）稳定;这些二聚体代表了衣壳形成的结构单元。然而，二硫键对于二聚化或组装不是必需的，因为Cys 61可以被Ala取代[9，10]而不影响任何过程。二聚体的界面显示出导致不平坦表面的突出尖峰。[6，13]虽然已经通过电子显微镜（EM）和X射线晶体学对HBV衣壳结构进行了广泛的结构研究[14]，但对其生物物理特性的了解有限。[15在这里，我们提供了来自大分子串联和离子迁移率质谱（MS）的数据[17-19]，这些数据与真空中HBV衣壳的稳定性和构象多样性有关。
The hepatitis B virus (HBV) is a major cause of liver disease in humans[1] and its non-infectious capsid is of interest for nanotechnology, including for drug-delivery applications. A precise biophysical characterization of these particles is of importance not only for these applications, but also because it may provide further insight into the replication cycle and assembly pathway of the virus, and thus contribute to the future development of drugs.[2,3] The HBV capsid protein (cp) forms icosahedral capsids of two sizes in vivo and in vitro (with triangulation numbers of T = 3 and T = 4[4] that contain 180 and 240 subunits, respectively[5–8]).The capsid protein has two domains—a core domain (amino acids 1–140) and a “protamine domain” (amino acids 150–183)—connected by a 10-residue linker;[9] of these, the core domain is necessary and sufficient for assembly of the capsid. The length of the linker and the conditions under which assembly take place determine the ratio of the T = 3 and T = 4 capsids obtained.[10] Capsid protein dimers are stabilized by an intermolecular four-helix bundle[11–13] and a disulfide bond within the bundle (Cys61); these dimers represent the building blocks for the formation of the capsid. However, the disulfide bond is not required for dimerization or assembly, as Cys61 can be replaced with Ala[9, 10] without affecting either process. The interfaces of the dimers display protruding spikes that result in an uneven surface.[6, 13] Although extensive structural studies of the HBV capsid structure have been performed by electron microscopy (EM) and X-ray crystallography,[14] knowledge of its biophysical properties is limited.[15, 16] Here, we present data from macromolecular tandem and ion mobility mass spectrometry (MS)[17–19] that have a bearing on the stability and conformational diversity of HBV capsids in vacuo.