Component Goal:
To fractionate native P. furiosus biomass by column chromatography in order to purify native multiprotein complexes. In collaboration with the other MAGGIE groups, these purified complexes will be identified by mass spectrometry (Fig 2C) and further elucidated structurally by SAXS. Ultimately the data derived here will allow successful expression of functional, recombinant multiprotein complexes in heterologous systems such as Escherichia coli and Sulfolobus solfataricus. These complexes will be analyzed for the number of co-purifying proteins by size exclusion chromatography combined with light scattering, and by native and denaturing gel analyses (Fig 2B), as well as for metal (Fig 2A) and organic cofactor content at UGA.
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