Development and Optimization of a Luciferin-Luciferase Activity Assay for the Detection of Nucleoside Diphosphate Kinase
AuthorSpeirs, Katie Marie
AdvisorBuxton, Iain L.O.
Biochemistry & Molecular Biology
Biochem and Molecular Biology
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Breast cancer is the most commonly occurring cancer among women. Metastasis of primary breast cancer tumors is the main cause of death related to the disease. Growing tumor cells require an adequate blood supply, and have an intrinsic ability to attract new blood vessels via a process called angiogenesis. Extracellular nucleosides are important to the growth of various cancers, as well as endothelial function and blood flow regulation. One factor secreted from breast cancer cells is nucleoside diphosphate kinase (NDPK), which functions to promote cell migration, growth, and tumor-mediated angiogenesis. Understanding the role of NDPK is important because identifying nucleoside function could lead to the prevention of metastasis development. The detection of NDPK activity in serum samples suggests that this may be one potential source for elevated ATP levels involved in purinergic mechanisms. The assay developed for the detection of NDPK utilizes the enzyme's transphosphorylation activity to generate ATP, which can be detected and quantified through luciferin-luciferase coupling.