Lee P M, Cherry R J, Bächi T
Virology. 1983 Jul 15;128(1):65-76. doi: 10.1016/0042-6822(83)90319-7.
The rotational mobility of Sendai virus glycoprotein spikes was measured by flash-induced transient dichroism of eosin triplet probes. The possible importance of this molecular motion for function was investigated by parallel assays of hemagglutination and fusion with erythrocytes. For mobility measurements, the glycoproteins were labeled on amino groups with eosin-5-isothiocyanate and on the galactose residues of the oligosaccharide chains with eosin-5-thiosemicarbazide. The decay of the absorption anisotropy of both probes, which has a time constant of about 100-200 musec at 37 degrees is attributed to the rotation of the proteins about an axis normal to the plane of the membrane. This motion was inhibited by crosslinking of the spike proteins with glutaraldehyde or by the specific binding of human erythrocyte glycophorin (a virus receptor) to the HN glycoprotein. Low values of the initial anisotropy for both probes indicate the existence of a second, faster motion. This is attributed to segmental motion of the glycoproteins. Segmental motion is inhibited by crosslinking with glutaraldehyde but appears to be little affected by interaction with glycophorin. The temperature dependence of the segmental and rotational motion of the proteins revealed a pronounced increase in mobility in the range of 30-35 degrees which was not paralleled by the lipid motion of the Sendai virus envelope membrane. Since the temperature dependence of virus-induced hemolysis has a similar characteristic, the mobility of glycoproteins appears to be correlated with the fusion activity. The hemagglutination activity, however, is not dependent on the mobility of the glycoprotein spikes.
通过嗜酸性三联体探针的闪光诱导瞬态二色性测量了仙台病毒糖蛋白刺突的旋转流动性。通过与红细胞的血凝和融合的平行测定，研究了这种分子运动对功能的可能重要性。为了进行流动性测量，用异硫氰酸荧光素对糖蛋白的氨基进行标记，并用硫代氨基脲荧光素对寡糖链的半乳糖残基进行标记。两种探针吸收各向异性的衰减，在37℃时时间常数约为100 - 200微秒，这归因于蛋白质围绕垂直于膜平面的轴的旋转。这种运动通过用戊二醛交联刺突蛋白或通过人红细胞血型糖蛋白（一种病毒受体）与HN糖蛋白的特异性结合而受到抑制。两种探针的初始各向异性低值表明存在第二种更快的运动。这归因于糖蛋白的片段运动。片段运动通过与戊二醛交联而受到抑制，但似乎受与血型糖蛋白相互作用的影响很小。蛋白质片段和旋转运动的温度依赖性显示，在30 - 35℃范围内流动性明显增加，这与仙台病毒包膜膜的脂质运动不同步。由于病毒诱导的溶血的温度依赖性具有类似的特征，糖蛋白的流动性似乎与融合活性相关。然而，血凝活性不依赖于糖蛋白刺突的流动性。
