Technology Bundle ID

Cell-based High-throughput High-content Assays Using Glycolytic Enzymes for Drug Discovery

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Lead Inventors
James Inglese (NCATS)
Patricia Dranchak (NCATS)
This technology includes an assay capable of monitoring glycosome formation for use in high throughput screening (HTS). The reversible assembly and disassembly of a multi-enzyme complex, known as the glycosome, visualized by GFP-labeled human phosphofructokinase-1 (PFK1), is employed as an intracellular marker in human cells to screen small molecule libraries under high-content imaging in a high-throughput fashion. The glycolytic enzymes have been proposed to form a multi-enzyme complex in the cell. It has recently been shown in several human cancer cells that human liver-type PFK1 forms cytoplasmic clusters by recruiting other rate-limiting enzymes in glycolysis and gluconeogenesis, indicating the formation of a multi-enzyme metabolic assembly, namely the “glucosome”. It appears that the enzymes in glucose metabolism are spatially organized into cytoplasmic clusters in human cells.
Commercial Applications
The assay may provide a means to discover agents targeting a novel aspect of cellular physiology important to diseases such as cancer.
Competitive Advantages
This assay enables a phenotypic analysis of regulatory aspects of cellular glycolysis that have are highly amenable to HTS.

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