A carboxy-terminal trimerization domain stabilizes conformational epitopes on the stalk domain of soluble recombinant hemagglutinin substrates.
A carboxy-terminal trimerization domain stabilizes conformational epitopes on the stalk domain of soluble recombinant hemagglutinin substrates.
复制标题
DOI：
10.1371/journal.pone.0043603
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Palese P
中科院分区：
文献类型：
--
作者：
Krammer F;Margine I;Tan GS;Pica N;Krause JC;Palese P
最近，发现了一类新的靶向病毒血凝素的茎域的广泛中和的抗流感病毒抗体。因此，这些新型抗体的诱导、分离、表征和定量已成为一个备受关注的研究领域。由于这些抗体中的大多数结合构象表位，因此用于检测和定量这些抗体的血凝素底物的结构完整性非常重要。在这里，我们评估这些抗体的结合，可溶性，分泌血凝素有或没有一个羧基末端三聚化结构域的基础上的天然三聚化结构域的T4噬菌体纤维蛋白。缺乏这样的结构域完全消除了与第1组血凝素的结合，并且还影响与第2组血凝素的结合。此外，三聚化结构域的存在积极影响可溶性血凝素在表达和纯化过程中的稳定性。我们的研究结果表明，羧基末端三聚化结构域是重组可溶性流感病毒血凝素茎表位结构完整性的必要条件。
Recently, a new class of broadly neutralizing anti-influenza virus antibodies that target the stalk domain of the viral hemagglutinin was discovered. As such, induction, isolation, characterization, and quantification of these novel antibodies has become an area of intense research and great interest. Since most of these antibodies bind to conformational epitopes, the structural integrity of hemagglutinin substrates for the detection and quantification of these antibodies is of high importance. Here we evaluate the binding of these antibodies to soluble, secreted hemagglutinins with or without a carboxy-terminal trimerization domain based on the natural trimerization domain of T4 phage fibritin. The lack of such a domain completely abolishes binding to group 1 hemagglutinins and also affects binding to group 2 hemagglutinins. Additionally, the presence of a trimerization domain positively influences soluble hemagglutinin stability during expression and purification. Our findings suggest that a carboxy-terminal trimerization domain is a necessary requirement for the structural integrity of stalk epitopes on recombinant soluble influenza virus hemagglutinin.
登录
查看更多内容
影响因子：
4.4
作者：
Cox MM;Patriarca PA;Treanor J
通讯作者：
Treanor J
影响因子：
4.7
作者：
Krammer, Florian;Nakowitsch, Sabine;Messner, Paul;Palmberger, Dieter;Ferko, Boris;Grabherr, Reingard
通讯作者：
Grabherr, Reingard
影响因子：
5.4
作者：
Steel, John;Lowen, Anice C.;Palese, Peter
通讯作者：
Palese, Peter
影响因子：
5.5
作者：
Khurana, Surender;Larkin, Christopher;Golding, Hana
通讯作者：
Golding, Hana
影响因子：
5.4
作者：
Krause, Jens C.;Tsibane, Tshidi;Crowe, James E., Jr.
通讯作者：
Crowe, James E., Jr.