Biological Sciences Division
Protein Function
In recognition of innovative separations technologies and mass spectrometry capabilities, the National Institutes of Health has established PNNL as a National Center for Research Resources in Proteomics. The Human Proteome Organization, an international consortium dedicated to the progress of proteomics, uses PNNL to provide expertise in technology and sample preparation. PNNL is also the proteomics lead for the Shewanella Federation, a consortium formed to characterize and model the biology of the metabolically versatile bacterium Shewanella oneidensis MR-1.
A library of yeast cells is incubated with fluorescently-labeled antigens and loaded into the BD FACS-ARIA cell sorter. Cells with antibodies capable of binding the antigen are visualized and isolated for further characterization.
Much of the real understanding of cellular systems and the roles of their constituents is based on proteomics, the study of protein expression patterns in organisms. The Protein Function group within the BSD has a four-tiered technological approach to proteomics. The first tier uses the Lab's High-Performance Mass Spectrometry Facility, which provides high sensitivity and measured mass accuracy and includes a novel accurate mass and time tag approach. The second tier is the use of an extensive library of single chain antibodies to generate affinity reagents for a variety of uses. The third tier of the Lab's approach involves high-throughput, high-sensitivity assay development to measure the activity of the proteins. The fourth tier applies the computational biology capability to interpret these data and build predictive models.
Staff in the Protein Function group use this four-tiered proteomics approach to study fundamental scientific questions such as the role of protein complexes in signal transduction and in microbial function, the process of DNA repair, the changes in cellular function that accompany malignant transformation, and the detoxification of metabolic products. Active research programs for practical applications of this unique proteomics approach include projects to define the components of normal human blood plasma, identify potential biomarkers for disease or exposure to toxic agents, produce new reagents for biodetection, and use nanomaterials to optimize the function of proteins and enzymes.
Based on the unique high-field mass spectrometry capabilities, laboratory automation, and bioinformatics developed at the W.R. Wiley Environmental Molecular Sciences Laboratory (EMSL), a Prototype Sample Processing and Proteomics Facility has been established at PNNL to support proteomic research in the following areas:
- protein production
- generation of affinity tags
- protein complex isolation and protein purification
- robotics prototyping
- protein analysis and complex characterization
- fundamental studies in protein-protein interactions.
Contact: Karin D. Rodland, Technical Leader