Active Enzyme Centrifugation of Amino Acid Decarboxylases

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This RUI project will test the hypothesis that different oligomers of multi-subunit enzymes with identical subunits may have different specific activities. Glutamate decarboxylase and arginine decarboxylase will be used. The sedimentation rate of the oligomers in the centrifugal field will be measured by active species followed by a spectrophotometric assay. The mass distribution of the enzyme will be followed in parallel band centrifugation experiments by absorption at 280 nm. The monomoer will be maintained in that form at the same total concentration as that of the largest oligomer by the nondenaturing by dissociating detergent, CHAPS. The sedimentation coefficient will permit identification of the oligomeric species present, with appropriate correction for the amount of bound detergent. The project should enhance our understanding of one aspect of the fundamental property of enzymes and at the same time offer excellent training to undergraduates on the theory and practice of ultracentrifugation.