Active Enzyme Centrifugation of Amino Acid Decarboxylases

本RUI项目将检验具有相同亚基的多亚基酶的不同低聚物可能具有不同的比活性的假设。将使用谷氨酸脱羧酶和精氨酸脱羧酶。低聚物在离心场中的沉降率将通过活性物质测量，然后进行分光光度测定。酶的质量分布将在平行带离心实验中通过280 nm的吸收来跟踪。单体将保持在与最大的低聚物相同的总浓度形式，通过解离洗涤剂CHAPS不变性。沉降系数将允许对存在的低聚物种类进行鉴定，并对结合洗涤剂的量进行适当的校正。该项目将增强我们对酶的基本性质的一个方面的认识，同时为本科生在超离心理论和实践方面提供良好的培训。

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.