Studies on the conformational structure and protective role of the measles virus envelope glycoproteins.

利用杆状病毒表达系统，将流感病毒血凝素(HA)蛋白的信号序列和后续两个残基(MAIIYLILLFTAVRG/DQ)连接到H蛋白胞外区的氨基末端，获得了基因控制的麻疹病毒血凝素(H)蛋白。重组H蛋白在Sf21细胞中得到高效表达，并能高效分泌到培养基中。重组H蛋白经抗麻疹抗体亲和层析和凝胶过滤层析纯化。通常，从1升培养物中获得1毫克纯化的H蛋白。重组H蛋白主要以单体形式存在。它具有抗单抗的抗原构象，并保留了受体结合活性，但缺乏HA活性。该方法可用于大规模分离组份疫苗的病毒糖蛋白，以及用于结晶学。

Genetically-manipulated measles virus hemagglutinin (H) protein was expressed via Baculovirus expression system, in which the signal sequence and the following two residues of the influenza virus hemagglutinin (HA) protein (MAIIYLILLFTAVRG/DQ) was attached to the amino-terminus of the ecto-domain of the H protein. The recombinant H protein was highly expressed in the Sf21 cells and it was efficiently secreted into the medium. Approximately 70% of the expressed protein was recovered in the medium.The recombinant H protein was purified through affinity-column using anti-measles antibody and the following gel-filtration column. Typically, 1 mg of the purified H protein was obtained starting from 1 liter of the culture. The majority of the recombinant H protein was present in a monomer. It possessed antigenic conformation against monoclonal antibodies and retained receptor binding activity but lacked HA activity. This protocol may be useful for large scale isolation of the viral glycoproteins for the component vaccines as well as for crystallography.