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Custom Vector Construction> | Specific siRNAs against PKM1 (50-GCGUGGAGGCUUCUUAUAA-30), PKM2 (50-CCAUAAUCGUCCUCACCAA-30) and SNAP-23 (50-CCAACAGAGAUCGU AUUGA-30) were purchased from RIBOBIO (Guangzhou, China). Recombinant SNAP-23 and expression vectors of wide type/mutant SNAP-23 were synthesized and purchased from Genscript (Nanjing, China). | Get A Quote |
Tumour cells secrete exosomes that are involved in the remodelling of the tumour-stromal environment and promoting malignancy. The mechanisms governing tumour exosome release, however, remain incompletely understood. Here we show that tumour cell exosomes secretion is controlled by pyruvate kinase type M2 (PKM2), which is upregulated and phosphorylated in tumours. During exosome secretion, phosphorylated PKM2 serves as a protein kinase to phosphorylate synaptosome-associated protein 23 (SNAP-23), which in turn enables the formation of the SNARE complex to allow exosomes release. Direct phosphorylation assay and mass spectrometry confirm that PKM2 phosphorylates SNAP-23 at Ser95. Ectopic expression of ... More