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

The ISCOMMM Team

Electron Cryo-microscopy and Image Analysis of Macromolecular Complexes

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

Cryo-EM Structure of Integrin α IIb β 3 Brian Adair and Mark Yeager

Docking of X-ray Structure of α V β 3 Ectodomain into Cryo-EM Map of α IIb β 3 Lower resolution EM maps of functionally important, transient states can be combined with high resolution X-ray structures to derive atomic models.

Integrin Molecular Models X-rayHigh affinityLow Affinity

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

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

The Scripps Research Institute and Scripps Clinic