TY  - JOUR
TI  - Determination of VPS34/PIK3C3 Activity in vitro Utilising 32P-γATP
AU  - Munson, Michael J.
AU  - Ganley, Ian G.
VL  - 6
IS  - 16
PY  - 2016
DA  - 2016/08/20
SP  - e1904
C1  - Bio-protocol 2016;6:e1904
DO  - 10.21769/BioProtoc.1904
UR  - https://doi.org/10.21769/BioProtoc.1904
AB  - VPS34 is the only class III phosphatidylinositol-3-kinase (PI3K) in mammalian cells and produces the vast majority of cellular phosphatidylinositol-3-phosphate [PI(3)P]. PI(3)P is a key signalling lipid that plays many membrane trafficking roles in processes such as endocytosis and autophagy. VPS34 is a key cellular regulator, loss of function can have catastrophic effects and is embryonic lethal (Zhou et al., 2011). The levels of cellular PI(3)P can be determined by fluorescent staining techniques and can be used to monitor effects upon VPS34 activity, however it is important to verify that any changes are mediated by VPS34, particularly as alternate pathways of PI(3)P production are possible such as via class II PI3Ks (Devereaux et al., 2013). Assaying VPS34 activity directly in vitro can be a key stage in delineating the action of a particular stimulus.
KW  - Phosphatidylinositol-3-phosphate
KW  - VPS34
KW  - PIK3C3
KW  - Lipid kinase assay
KW  - TLC
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False