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Structural Determination of Protein_Lab Meeting.pptx
- 1. Structural Determination of Protein: a brief introduction
- 2. Why do weneed wet-lab experimental data? Protein modelling software programs (AlphaFold, Tasser, SWISSModel, etc) are not without limitations Understanding the enzymatic mechanisms Elucidating protein-protein/protein-ligand interactions Solving novel protein structures Techniques for structural determination X-ray Crystallography Cryo-Electron Microscopy (Cryo-EM) Nuclear Magnetic Resonance (NMR) https://peakproteins.com/a-comparison-of-the-structural-techniques-used-at-sygnature-di x-ray-crystallography-nmr-and-cryo-em/#
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- 7. Cryo-EM Workflow Dhakal etal. (2023) Scientific Data
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- 10. Choosing the besttechnique https://www.criver.com/resources/x-ray-crystallography-or-cryo-em-which-solution-should-you-choose
- 11. X-Ray Crystallography vsCryoEM X-Ray Crystallography Advantages • Well established • No size limitation • Can achieve atomic resolution of structure in crystalline state CryoEM Disadvantages • Need to make crystal • Crystals need to be in the right order • Limited types of sample > membrane proteins and large macromolecules are difficult to crystallise Advantages • Can achieve near atomic resolution of sample in the native state • Can be used for large macromolecules • Multiple states of macromolecule can be purified in silico from a single data set Disadvantages • It is difficult to image small proteins • Needs a large computational power (storage, GPU) • Slow data acquisition, processing, and analysis