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Hierarchy of Biological Complexity

Hierarchy of Biological Complexity. LIFE. Interactions of machines (molecular and cellular dynamics). Macromolecular machines. Proteins and nucleic acids. Sequences. GENOMES. The ISCOMMM Team. Electron Cryo-microscopy and Image Analysis of Macromolecular Complexes. Experimental Design.

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Hierarchy of Biological Complexity

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  1. Hierarchy of Biological Complexity LIFE Interactions of machines (molecular and cellular dynamics) Macromolecular machines Proteins and nucleic acids Sequences GENOMES

  2. The ISCOMMM Team

  3. Electron Cryo-microscopy and Image Analysisof Macromolecular Complexes

  4. Experimental Design • Large-scale production of purified vaccinia virus particles • Isolation and purification of vaccinia molecular machines • Identification of protein composition by mass spectrometry • Generation of antibodies directed again proteins in the machine • Formation of antibody-molecular machine complexes • Preparation of frozen-hydrated specimens • Automated recording of cryo-EM images • Automated extraction of particle images from electron micrographs • Automated determination of 3D density maps • Protein localization by use of antibody labeling and difference map analysis • Validation of computational predictions of protein-protein interactions

  5. Cryo-EM Structure of Integrin αIIbβ3 Brian Adair and Mark Yeager

  6. Docking of X-ray Structure of αVβ3 Ectodomaininto Cryo-EM Map of αIIbβ3 Lower resolution EM maps of functionally important, transient states can be combined with high resolution X-ray structures to derive atomic models.

  7. Integrin Molecular Models X-ray High affinity Low Affinity

  8. Cryo-EM density fitting Protein-protein docking prediction & density correlation fitting Visual fitting

  9. Cryo-EM density fitting Protein-protein docking prediction & density correlation fitting Visual fitting

  10. The Scripps Research Institute and Scripps Clinic

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