GTL Facilities Characterization and Imaging of Molecular Machines Lee Makowski.
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Transcript of GTL Facilities Characterization and Imaging of Molecular Machines Lee Makowski.
GTL FacilitiesCharacterization and Imaging of
Molecular Machines
Lee Makowski
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Genomes to Life Facilities for 21st Century Biology
Facility III Characterization and Imaging of
Molecular Machines
Isolate the repertoire of molecular machines.
Characterize machines in terms of composition and molecular organization.
Facility III
Facility I
Facility IV
Facility II
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Nearly every major process in a cell is carried out by assemblies of 10 or more protein molecules
Identifying the complexes and understanding their function represents a huge challenge
Facility 3: Characterization and Imaging of Molecular Machines
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Facility 3: Characterization and Imaging of Molecular Machines
GOAL: Isolation, identification and characterization of 1000’s of molecular machines; providing revolutionary new information and capabilities to the biological community
Repertoire of protein complexes
Biophysically characterize complexes
Develop principles, theory and predictive models
Understanding how molecular machines operate at the molecular level will unlock the capability to control useful biochemical processes in a microbe and apply them to DOE mission needs.
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Will Require State-of-the-Art Analytical and Computational Tools
Automated molecular machine stablization/isolation techniques
High performance mass spectrometry
Scattering techniques—optical, neutron, x-ray
Imaging tools-cryoEM, optical, force microscopy
Bioinformatics tools and databases/data analysis tools
Modeling and simulation
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Identifying the parts of a molecular machine
Is the first step to understanding
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Determining the relative positions of gene products in a machine can provide
important clues to their functions
Yeast spindle pole body - microtubule organizing center
Novel imaging tools will be needed to characterize the relative positions of each one
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Detailed images will be needed to complete the linkage of structure to function
Zhou et al., Science 2000
300 keV cryoEM
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Interaction Mapping and Imaging are Computationally Intensive
For each organism:
•thousands of gene products
•tens of thousands of intermolecular interactions
For each complex:
• thousands of images
• studied under different conditions
• localize each component
• data in three dimensions
… and untold numbers of molecular complexes
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Developing models for the assembly, disassembly and function of molecular machines will be
computationally challenging
Machines
Proteins
Cellular systems
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Facility IV Cellular Systems
Gene Sequence
Gene Sequence
High Validity DataResource for
Biological Community
High Validity DataResource for
Biological Community
Facility III: Characterization and Imaging of Molecular Machines
Facility IIProteome
Data
Facility IReagents
Microbe Growth
Complex Isolation
Data Interpretation/
Archival, Modeling
Image Complexes
BiophysicalCharacterization
ComponentIdentification