Technology Bundle ID
TAB-3543

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

Applications
Linked ID
E-178-2018-0
Lead Inventors
James Inglese (NCATS)
Co-Inventors
Patricia Dranchak (NCATS)
ICs
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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