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Published on: June 28, 2013
Crystallization and preliminary X-ray diffraction analysis of central structure domains from mumps virus F protein
Yueyong Liu1, Yanhui Xu, Jieqing Zhu
1Department of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China.
Abstract:
Fusion of members of the Paramyxoviridae family involves two glycoproteins: the attachment protein and the fusion protein. Changes in the fusion-protein conformation were caused by binding of the attachment protein to the cellular receptor. In the membrane-fusion process, two highly conserved heptad-repeat (HR) regions, HR1 and HR2, are believed to form a stable six-helix coiled-coil bundle. However, no crystal structure has yet been determined for this state in the mumps virus (MuV, a member of the Paramyxoviridae family). In this study, a single-chain protein consisting of two HR regions connected by a flexible amino-acid linker (named 2-Helix) was expressed, purified and crystallized by the hanging-drop vapour-diffusion method. A complete X-ray data set was obtained in-house to 2.2 A resolution from a single crystal. The crystal belongs to space group C2, with unit-cell parameters a = 161.2, b = 60.8, c = 40.1 A, beta = 98.4 degrees. The crystal structure will help in understanding the molecular mechanism of Paramyxoviridae family membrane fusion.
Insights
Researchers determined the crystal structure of a key mumps virus fusion protein component. This finding advances understanding of how Paramyxoviridae viruses fuse with host cell membranes.
Area of Science:
- Virology
- Structural Biology
- Biochemistry
Background:
- Paramyxoviridae family viruses, including mumps virus (MuV), utilize attachment and fusion glycoproteins for cell entry.
- Viral fusion involves conformational changes in the fusion protein, driven by receptor binding.
- Two conserved heptad-repeat (HR) regions (HR1 and HR2) are hypothesized to form a six-helix bundle during fusion, but its structure in MuV remains unknown.
Purpose of the Study:
- To determine the crystal structure of the MuV fusion protein's HR regions in a post-fusion conformation.
- To provide insights into the molecular mechanisms underlying membrane fusion in the Paramyxoviridae family.
Main Methods:
- Expression and purification of a single-chain protein construct containing both HR1 and HR2 regions linked by a flexible peptide.
- Crystallization of the 2-Helix protein using the hanging-drop vapour-diffusion method.
- X-ray diffraction data collection to 2.2 Å resolution.
Main Results:
- A single crystal of the 2-Helix protein was obtained, belonging to space group C2.
- Unit-cell parameters were determined: a = 161.2 Å, b = 60.8 Å, c = 40.1 Å, β = 98.4°.
- A complete X-ray data set was collected, enabling structural determination.
Conclusions:
- The determined crystal structure provides a high-resolution view of the MuV fusion protein's HR regions.
- This structural information is crucial for understanding the molecular basis of membrane fusion in Paramyxoviridae viruses.
- The findings lay the groundwork for developing antiviral strategies targeting viral fusion.
