Dranchak, Patricia (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.
The assay may provide a means to discover agents targeting a novel aspect of cellular physiology important to diseases such as cancer.
This assay enables a phenotypic analysis of regulatory aspects of cellular glycolysis that have are highly amenable to HTS.