RESEARCH

Specific Aims

1.  Examine the roles of active site residues in Cam.

Crystal structures (8) reveal E62, Q75, N73, N202, F140, and F132 as possible residues involved in CO₂ hydration; however, the exact roles of these residues are uncertain.  Kinetic analyses and crystal structures of replacement variants will be used to investigate the roles of these residues.  These data will contribute to a better understanding of Cam active site dynamics.

2. Determine the physiological function of Cam in Methanosarcina thermophila.

The physiological function of Cam has not been investigated although hypotheses have been advanced.  The cellular location of Cam is paramount to addressing these hypotheses and is a primary goal of this aim.  Discovering Cam’s physiological function may expose a novel role for CA’s in anaerobic methanogens and provide a better understanding of acetate metabolism in M. thermophila.

 3. Confirm and characterize putative g-class CA’s.           

Cam is the only characterized g-class CA in spite of a wide distribution of putative g-class CA’s in all domains of life.  Sequence alignments reveal putative g-class CA’s from anaerobic methanogens Methanobacterium thermoautotrophicum (Cam-1) and Methanocaldococcus jannaschi (Cam-2).  Characterizations these putative g-class CA’s will contribute to general understanding of the class and increase comprehension of Cam’s mechanistic and physiological qualities.

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