The Presence and Cleavage of Interpeptide Disulfide: Bonds in Viral Glycoproteins1
The Presence and Cleavage of Interpeptide Disulfide: Bonds in Viral Glycoproteins1
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肽间二硫化物的存在和裂解：病毒糖蛋白中的键1
DOI：
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发表时间：
1979
期刊：
影响因子：
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通讯作者：
Y. Okada
中科院分区：
文献类型：
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作者：
M. Ozawa;A. Asano;Y. Okada
摘要本文描述了仙台病毒（HVJ） Aemagglutinin and neuroa -minidase)糖蛋白HANA和F （fusion）糖蛋白中肽间二硫键的存在和性质。在HANA的情况下，相同或非常相似分子量的亚基通过二硫键相互连接。通过添加1mm的二硫苏糖醇可以很容易地实现键的切割，同时失去糖蛋白的生物活性。在连接键断裂后，所有的HANA蛋白亚基仍然结合在病毒膜上。为了观察F糖蛋白F1和F2亚基之间肽间二硫键的断裂，需要比HANA蛋白更高浓度的巯基化合物。在变性或非变性条件下，二硫键的分裂都不能将F蛋白的两个片段从病毒粒子中释放出来。因此，F糖蛋白似乎至少有两个膜结合位点，一个在F1上，另一个在F2上。另一方面，连接流感病毒HA1和HA2亚基的二硫键在非变性条件下几乎不会断裂。加入8 M尿素或6 M胍HCl，可以完全灭活HA活性，这是由硫醇化合物分裂二硫键所必需的。有趣的是，HA1提交在切割后从病毒粒子中释放出来。因此，与HVJ的F1和F2不同，HA1亚基似乎与膜没有疏水结合位点。提出了这些亚基在病毒膜上排列的模型。
Abstract The presence and nature of interpeptide disulfide bonds in HANA (Aemagglutinin and neura-minidase) glycoprotein and F (fusion) glycoprotein of HVJ (Sendai virus) are described. In the case of HANA, subunits of the same or very similar molecular weight were inter connected with a disulfide bond(s). Cleavage of the bond(s) can easily be achieved by the addition of 1 mM dithiothreitol with concomitant loss of the biological activities of the glyco-protein. After splitting of the interconnecting bonds, all the HANA protein subunits remained bound on the viral membrane. To observe the cleavage of the interpeptide disulfide bond between the F1 and F2 subunits of F glycoprotein, higher concentrations of sulfhydryl compounds were required than were necessary for HANA protein. Splitting of the disulfide bond under either denaturing or non-denaturing conditions failed to release both segments of F protein from the virion. Therefore, F glycoprotein seems to have at least two membrane binding sites, one on F1 and the other on F2. On the other hand, the disulfide bond which connects the HA1 and HA2 subunits of influenza virus is hardly cleaved under non-denaturing conditions. Addition of 8 M urea or 6 M guanidine HCl, which completely inactivates HA activity, was necessary for the splitting of this disulfide bond by thiol compounds. Interestingly, the HA1 submit was released from the virion after the cleavage. Thus, unlike F1 and F2 of HVJ, the HA1 subunit seems to have no hydrophobic binding site to the membrane. A model for the arrangement of these subunits on the viral membrane is proposed.