| Accession | PRJCA015948 | ||||||||||||||||||||||||||||||||
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| Title | Cryo-EM in situ structural dynamic analysis technology | ||||||||||||||||||||||||||||||||
| Relevance | Medical | ||||||||||||||||||||||||||||||||
| Data types |
Map
Models of biological macromolecular structures |
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| Organisms |
Severe acute respiratory syndrome coronavirus 2
enterovirus D68 Mus musculus influenza A virus Chlamydomonas reinhardtii Bos taurus Coxsackievirus A16 |
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| Description | Many macromolecular complexes and related structural forms of large cellular ultrastructures (e.g cilia) may exist only in their native environment, and may also contain information such as transient binding mates and conformations in specific environments. Cryo-electron tomography (cryo-ET) preserves the complete network of conformations and interactions of each molecule by imaging undisturbed cells, becoming a central means of providing a complete structural description of the cell's "native molecular landscape". However, due to the excessive thickness of the cell sample, the tilt and movement of the sample, the low signal-to-noise ratio/contrast caused by the low electron dose, and the problem of missing cones, the current structural analysis using cryo-ET combined with sub-tomogram average method is usually difficult to break through the resolution of 10 angstroms, and the dynamic structure study is also very limited. Constraints include: macromolecular complexes or cell ultrastructure are difficult to purify, frozen samples are fragile, cell thinning is not easy to achieve, and target macromolecules are difficult to accurately locate; The image data contrast is extremely low and the high-resolution information is missing, resulting in the average accuracy and speed of sub-tomogram is not high, which requires a lot of computing resources. Underfocus determination and CTF correction are challenging; A large number of cryo-ET dumping series data collection time-consuming, etc. In recent years, the development of VPP phase plate technology, direct electron detection technology, spherical aberration correction technology, cell/tissue thinning technology and sub-tomogram average calculation using cryo-FIB has laid a foundation for breaking through the above limiting factors and realizing the three-dimensional reconstruction and dynamic structure analysis of macromolecular complexes with in-situ sub-nanometer resolution.Relying on the cryo-EM facility of the National Center for Protein Science (Shanghai), the applicant will explore the new high-resolution, high-contrast imaging technology of cryo-ET, develop the related sub-tomography averaging and classification technology, study affinity grid sample preparation technology and subunit localization technology, take cilia and the key macromolecular complexes contained therein (such as IFT and radiation axis RS complex) as the research object, and complete the establishment of cryo-EM in situ structure dynamic analysis technology for macromolecular complexes. It was also applied to the study of the in situ dynamic structure of important macromolecular complexes in project 1-3. | ||||||||||||||||||||||||||||||||
| Sample scope | Synthetic | ||||||||||||||||||||||||||||||||
| Release date | 2023-03-30 | ||||||||||||||||||||||||||||||||
| Grants |
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| Submitter | Yao Cong (conglab@sibcb.ac.cn) | ||||||||||||||||||||||||||||||||
| Organization | Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences | ||||||||||||||||||||||||||||||||
| Submission date | 2023-03-29 |
| Resource name | Description |
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