Workshop Structural Proteomics of Biological Complexes

Questions Why is it timely to study complexes ? What are the most appropriate model organisms ? How to predict relevant complexes ? How to purify complexes ? How to validate their biological functions ? What are the necessary technologies ? What are the realistic goals and timetables ? What are the necessary resources to accomplish the goals ? What are the funding mechanisms for such an initiative ?

Workshop Agenda Session I Cell Biology and Complexes Session II Biological Processes and Protein Machines Session IIIGenetic and Chemical Approaches Session IV Computational Approaches Session V Electron Cryomicroscopy Session VI Crystallography Session VII Proteomics Centers Session VIIIRecommendations

Baylor College of Medicine Wah Chiu

Electron Cryomicroscopy: A Structural Biology Tool Instrument resolution beyond 2.2 Å

Single machines: 9 – 6.8 Å  helices and ß sheets visualized Structural Features Revealed by Electron Cryomicroscopy Crystalline arrays : Å polypeptide traced Helical arrays: Å  helices and ß strands resolved Subcellular assemblies and whole cell: Å identify components and domains Single machines: 9 – 6.8 Å  helices and ß sheets visualized Single machines: 9 – 6.8 Å  helices and ß sheets visualized

GroEL D Chen

9 Å Structure of GroEL S Ludtke

9 Å Structure of GroEL

6.8 Å Structure of Rice Dwarf Virus Zhou et al. Nature SB (2001) 8:868-73

Rice Dwarf Virus Outer Shell Protein P8

Pseudo Atomic Model of RDV

Images of P22 Phage 500Å Procapsid Mature phage

Structural Transitions in Phage Maturation Jiang et al (2003) Nature SB 10:131-5

Issues in CryoEM for Studying Single Particles of Complexes High structural purity Chemical vs computational High resolution Now 7-9 Å Future 3-4 Å High throughput analysis Now 5-10 months Future 1- few weeks

Experimental and Computational Processes Biochemical Purification Cryo-Specimen Preparation Data Collection Data Digitization Pre-processing Initial Model Structure Refinement Structure Visualization Structure Analysis Biochemical Purification Cryo-Specimen Preparation Data Collection Data Digitization Pre-processing Initial Model Structure Refinement Structure Visualization Structure Analysis

Manpower Chemists or Biochemists Physicists or Engineers Computational Scientists New Instrumentation EM CCD Camera Freezing Apparatus High Performance Computers Resources Needed