Crystallization of biologically active hemagglutinin-neuraminidase glycoprotein dimers proteolytically cleaved from human parainfluenza virus type 1.
Crystallization of biologically active hemagglutinin-neuraminidase glycoprotein dimers proteolytically cleaved from human parainfluenza virus type 1.
复制标题
从人副流感病毒 1 型中蛋白水解裂解出具有生物活性的血凝素-神经氨酸酶糖蛋白二聚体的结晶。
DOI：
10.1128/jvi.66.12.7597-7600.1992
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发表时间：
1992
影响因子：
5.4
通讯作者：
Portner,A
中科院分区：
文献类型：
--
作者：
Takimoto,T;Laver,WG;Murti,KG;Portner,A
我们分离，纯化，并确定了人副流感病毒1型的血凝素-神经氨酸酶（HN）的特征，最终目标是生产适合三维X射线结构分析的晶体。链霉蛋白酶用于直接从病毒颗粒切割HN分子的球状头部，形成HN单体和二聚体。纯化的二聚体保留了神经氨酸酶和血细胞吸附活性，并被14种抗HN单克隆抗体识别，证明HN抗原结构和功能完整。二聚体的N-末端序列分析显示，在氨基酸136或137处发生切割，释放C-末端438或439个氨基酸。在电子显微镜下，二聚体呈现为两个盒状结构，每个约5 × 5 nm。当纯化的HN二聚体通过对20%聚乙二醇3350的蒸汽扩散以悬滴形式结晶时，它们形成矩形板和针状晶体。矩形晶体衍射X射线，表明有序的原子结构。然而，分辨率约为10 A（1 nm），不足以进行三维结构分析。通过增加晶体的尺寸和质量来提高分辨率的实验正在进行中。
We isolated, purified, and characterized the hemagglutinin-neuraminidase (HN) of human parainfluenza virus type 1, with the ultimate goal of producing crystals suitable for three-dimensional X-ray structure analysis. Pronase was used to cleave the globular head of the HN molecule directly from virus particles, forming HN monomers and dimers. The purified dimers retained neuraminidase and hemadsorption activity and were recognized by 14 anti-HN monoclonal antibodies, demonstrating intact HN antigenic structure and function. N-terminal sequence analysis of the dimers showed that cleavage had occurred at amino acid 136 or 137, freeing the C-terminal 438 or 439 amino acids. On electron micrography, the dimer appeared as two box-shaped structures, each approximately 5 by 5 nm. When the purified HN dimers were crystallized in hanging drops by vapor diffusion against 20% polyethylene glycol 3350, they formed both rectangular plates and needlelike crystals. The rectangular crystals diffracted X-rays, indicating an ordered atomic structure. However, the resolution was approximately 10 A (1 nm), insufficient for three-dimensional structural analysis. Experiments to improve the resolution by increasing the size and quality of the crystals are in progress.