Synthesis and Structural Probing of Protein Networks in Vitamin B6 Biosynthesis.
EMSL Project ID
50165
Abstract
We propose to develop a multi-modal experimental pipeline, integrating multiple existing and emerging EMSL capabilities, including cell-free protein expression, Native Mass Spectrometry (Native MS), Electron Paramagnetic Resonance (EPR) and Cryo-Electron Microscopy (Cryo-EM). The combination of these tools will provide functional insights into hetero-association of various protein multiunit complexes that have proved intractable by other means. To test these techniques in a short time frame, the protein regulation network of vitamin B6 production in plants, namely PDX protein family, is chosen as a pilot system. The scientific scope and combinatorial approach is in line with the focus of FY18 Satellite project for the Subcellular Thrust in the area of "Expression and structural analysis of proteins in pathways of bioenergy relevance or signaling". Upon refinement, this experimental strategy will be a valuable tool that can be easily applied to any organism and pathway of interest.
Project Details
Start Date
2018-04-04
End Date
2018-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Co-Investigator(s)
Team Members
Related Publications
Novikova, I., Zhou, M., Shaw, J., Hellmann, H., & Evans, J. (2019). Dual Native MS and Cryo-EM Approach to Resolve Heteromeric Protein Assemblies and Subunit Stoichiometry. Microscopy and Microanalysis, 25(S2), 1214-1215. doi:10.1017/S1431927619006809