NMR and EPR Structural Characterization of c-Type Cytochromes and their Complexes from Contaminant Metal Remediating Iron-Respiring Bacteria
EMSL Project ID
26709
Abstract
Abstract: While significant progress has been made regarding the genetics and genomics of dissimilatory metal-transforming bacteria and the redox proteins they possess, very little has yet been done to elucidate structural information of these proteins. This lack of progress is the result of 1) the great difficulty of purification and manipulation of these proteins, and 2) the lack of success of crystallization of these proteins for subsequent study. The Magnuson Microbiology Group at Idaho State University has surmounted some of these obstacles, and we have produced several proteins in sufficient quantity for biochemical analyses. Thus, we are uniquely poised to partner with the EMSL to apply high-resolution NMR and EPR techniques towards the elucidation of structure and function of these very interesting but poorly understood proteins. Our group will purify selected cytochromes c from two distinct model organisms (Acidiphilium cryptum and Geobacter sulfurreducens) of metals remediation importance, and submit these proteins for NMR and EPR analyses at the EMSL, using both medium and high resolution instruments. High-resolution NMR can reveal structural information for these membrane associated proteins without the need for crystallization, and can also be used for examination of protein-protein complexes that play a role in electron transfer to Fe(III) and contaminant metals such as uranium. EPR spectroscopy can provide information regarding the spin states of iron atoms within the protein, which are central to the redox functionality of these cytochromes. Our work will thus contribute directly to EMSLs mission in the environmental molecular sciences, as well as the EMSL Science Theme of Biological Interactions and Dynamics. Our past relationship and User Agreement with the Electron Microscopy Capability at EMSL has resulted in 2 manuscripts, so we are confident that an NMR-based project would be equally productive.
Project Details
Project type
Exploratory Research
Start Date
2007-10-01
End Date
2008-10-05
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Cort JR, M Swenson, and TS Magnuson. 2011. "1H, 13C, and 15N backbone, side-chain, and heme chemical shift assignments for oxidized and reduced forms of the monoheme c-type cytochrome ApcA isolated from the acidophilic metal-reducing bacterium Acidiphilium cryptum." Biomolecular NMR Assignments 5(1):89-92. doi:10.1007/s12104-010-9274-1