GTL User Facilities

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GTL User Facilities

• Facility II
• Whole Proteome Analysis

Michelle V. Buchanan
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Genomes to Life User Facilities for 21st Century
Biology

• Facility II Whole Proteome Analysis
• Measure proteome and metabolites under
  well-defined conditions
• Gain first insight into function of proteins
• what pathways and processes are present under
  what conditions
• regulatory network structure and connectivity for
  individual processes

Facility II
Facility I
Facility IV
Facility III
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GTL Facility II Whole Proteome Analysis

• Key capabilities will include
• Growth of organisms under controlled conditions
• High-throughput approaches for sample preparation
  prior to analysis.
• High throughput techniques for the identification
  and quantification of proteins, metabolites.
• New computational tools for interpretation and
  modeling whole proteome data
• Databases and tools for interpreting, archiving,
  and disseminating data and models to the greater
  biological community.

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Genome-scale, comprehensive determination of
microbial proteomes will require high throughput
approaches for sample preparation and analysis

• Novel cell cultivation
• On-line analysis of metabolites
• Robotics and automation
• Chip expression assays

• Microsample handling
• Single cell analysis
• Imaging
• Computation and Informatics

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Deinococcus radiodurans Proteome

• BER pilot project developed MS-based approach for
  proteome analysis using FTICR and accurate mass
  tags (AMTs)
• Permits proteome to be rapidly identified
• Automated data collection and interpretation

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Protein Analysis of Whole Cells on Microfluidic
Devices
J.M. Ramsey, et al
Note arrows depict direction of flow.
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Impact of Facility II

• High fidelity data openly accessible to enable
  scientific studies
• Economy of scale
• Critical base information for Facilities III and
  IV
• Proteome database
• Models of pathways, processes and regulatory
  networks
• Empower the general microbiological and
  biological communities
•  

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Facility II Whole Proteome Analysis
Gene Sequence
Protein Processing
Mass Spectrometry
Microbe Growth
Data Interpretation/ Archival, modeling
Metabolite Analysis
Validation
High validity data resource for biological
community
Facility III Complexes
Facility IV Cellular Systems
Facility I Reagents