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Updated: May 29, 2026
Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques
Published on: July 5, 2018
Crystallization strategy for the glycoprotein-receptor complex between measles virus hemagglutinin and its cellular
Takao Hashiguchi1, Toyoyuki Ose, Marie Kubota
1Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060- 0812, Japan.
Abstract:
Measles virus (MV), one of the most contagious agents, infects immune cells using the signaling lymphocyte activation molecule (SLAM) on the cell surface. A complex of SLAM and the attachment protein, hemagglutinin (MVH), has remained elusive due to the intrinsic handling difficulty including glycosylation. Furthermore, crystals obtained of this complex are either nondiffracting or poorly-diffracting. To solve this problem, we designed a systematic approach using a combination of the following techniques; (1) a transient expression system in HEK293SGnTI(-) cells, (2) lysine methylation, (3) structure-guided mutagenesis directed at better crystal packing, (4) Endo H treatment, (5) single-chain formation for stable complex, and (6) floating-drop vapor diffusion. Using our approach, the receptor-binding head domain of MV-H covalently fused with SLAM was successfully crystallized and diffraction was improved from 4.5 Å to a final resolution of 3.15 Å . These combinational methods would be useful as crystallization strategies for complexes of glycoproteins and their receptors.
Insights
Researchers developed a novel method to crystallize the measles virus hemagglutinin (MV-H) and signaling lymphocyte activation molecule (SLAM) complex. This breakthrough improves understanding of viral entry mechanisms.
Area of Science:
- Virology
- Structural Biology
- Biochemistry
Background:
- Measles virus (MV) is highly contagious, infecting immune cells via the signaling lymphocyte activation molecule (SLAM).
- The complex between MV's hemagglutinin (MV-H) and SLAM has been difficult to study due to handling challenges and poor crystal quality.
Purpose of the Study:
- To develop a robust method for crystallizing the MV-H and SLAM complex.
- To achieve high-resolution structural data of the MV-H/SLAM interaction.
Main Methods:
- A multi-pronged approach including transient expression in HEK293SGnTI(-) cells, lysine methylation, and structure-guided mutagenesis.
- Endo H treatment and single-chain formation were employed to stabilize the complex.
- Floating-drop vapor diffusion was used for crystallization.
Main Results:
- Successful crystallization of the receptor-binding head domain of MV-H covalently fused with SLAM.
- Improved diffraction from 4.5 Å to a resolution of 3.15 Å.
Conclusions:
- The developed combinational strategy is effective for crystallizing challenging glycoprotein-receptor complexes.
- This method provides a pathway to structural insights into measles virus infection.
