Luminometer for the discovery of novel bioactive compounds

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Calmodulin (CaM)-dependent 3',5'-cyclic nucleotide phosphodiesterase (CaM-PDE) is a cellular enzyme that hydrolyzes cyclic adenosine monophosphate (cAMP) to form 5'-adenosine monophosphate. In normal human cells, elevated levels of cAMP is required to inhibit excessive proliferation and to induce normal cell death. However, in diseased states such as cancer, there is excessive levels of CaM-PDE, which decreases cAMP concentrations and disrupts the normal cell death that is required to check excessive cellular proliferation. Current evidence also indicates that excessive levels of CaM-PDE play significant roles in several pathobiologic processes leading to kidney fibrosis and hypertrophy of the heart. The aim of this proposal is to purchase a luminometer that will enhance discovery and identification of CaM-binding peptides that inhibit the activity of CaM-PDE with potential application as therapeutic agents against chronic diseases such as cancer and kidney fibrosis. The luminometer is the most sensitive and accurate instrument for measuring the light output during CaM-PDE reaction that has been coupled to the luciferin-luciferase system. The instrument also provides for the most rapid form of analysis of CaM-PDE activity. Therefore, the instrument will enhance the screening of several hundreds of potential peptide inhibitors with high accuracy and reliability over a short period of time. However, there is no luminometer in the Department of Human Nutritional Sciences at the University of Manitoba. Results provided by the instrument will identify bioactive peptides that can be used to formulate functional foods and nutraceuticals, in addition to providing fundamental information on the structure and activity relationships of the CaM-binding peptides. Technology developed from this work will facilitate value-added utilization of Canadian-grown crops such as peas, flaxseed, soybean with economic returns to the Canadian economy. Availability of the instrument will enhance interdisciplinary training of highly qualified personnel such as postdoctoral fellows, graduate and undergraduate students in the areas of food science and life sciences related to human health.