# Presentation: 117259

## GTL Facilities

***Characterization and Imaging of Molecular Machines***

**Lee Makowski**

## 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**** **

## Facility 3: Characterization and Imaging of Molecular Machines

***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***

## 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. ***** **

## 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

## Identifying the parts of a molecular machine 

***Identifying the parts of a molecular machine ***

***Is the first step to understanding ***

## Determining the relative positions of gene products in a machine can provide important clues to their functions

***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***

## Detailed images will be needed to complete the linkage of structure to function

**Detailed images will be needed to complete the linkage of structure to function**

- Zhou et al., Science 2000

- 300 keV cryoEM

## Interaction Mapping and Imaging are Computationally Intensive

**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**

## Developing models for the assembly, disassembly and function of molecular machines will be computationally challenging 

***Machines***

***Proteins***

***Cellular systems***

## Facility III:  Characterization and Imaging of Molecular Machines

**   ****Facility IV ****Cellular **
**Systems**

**Gene Sequence**

**High Validity Data**
**Resource for **
**Biological Community**

**Facility III:  ****Characterization and Imaging of Molecular Machines**

**Facility II**
**Proteome**
** ****Data**

**Facility I**
**Reagents**