Homogeneous sugar modification improves crystallization of measles virus hemagglutinin

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• 作者：Takao Hashiguchi ; Mizuho Kajikawa ; Nobuo Maita ; Makoto Takeda ; Kimiko Kuroki ; Kaori Sasaki ; Daisuke Kohda ; Yusuke Yanagi ; Katsumi Maenaka
• 关键词：Measles virus ; Hemagglutinin ; Crystallization ; Restricted N-glycosylation ; HEK293S-GnTI(&#x2212 ; ) cells ; HEK293T cells
• 刊名：Journal of Virological Methods
• 出版年：2008
• 期刊代码：180_01660934
• 类别：med
• 出版时间：April 2008
• 卷：149
• 期：1
• 页码：171-174
• 文件大小：395 K

摘要

Measles virus (MV) enters cells by binding to the signaling lymphocyte activation molecule (also called CD150) on the cell surface, and thus shows the lymphotropism and immunosuppressive effects. The head domain (residues Asp¹⁴⁹ to Arg⁶¹⁷) of the MV hemagglutinin (MV-H), the attachment protein, was produced using a transient expression system in HEK293T cells. The purified MV-H protein was heterogeneous because of a variety of complex-sugar modifications. The complex-sugar-type MV-H was crystallized successfully, and the crystals belonged to the space group P41212 with the unit cell dimension of a = b = 134 Å, c = 100 Å, but diffracted only to 3.0 Å resolution. MV-H was also expressed in HEK293SGnTI(−) cells lacking the N-acetylglucosaminyltransferase I activity, which render N-linked glycans of the proteins restricted and homogeneous, producing the oligomannose, Man₅GlcNAc₂. The native and selenomethionyl derivative proteins of the oligomannose-type MV-H were crystallized, and the native crystals well diffracted to 2.6 Å resolution. Thus, homogeneous sugar modification may be useful for improved crystallization of heavily sugar-modified viral envelope proteins.