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Analysis of PtdInsP Kinase Activity of PIP5K3 in Vitro.
Purified GST-PIP5K3 protein or GST alone was tested for PtdInsP kinase activity on different lipid mixtures containing no lipids (−), synthetic diC16:0 PtdIns4P, synthetic diC16:0 PtdIns5P, or natural PtdIns4P (from brain; nPtdIns4P) in the presence of [γ-32P]ATP. As positive controls, mammalian type I (mPIP5K) or type II (mPIP4K) PtdInsP kinases were used. The position of PtdIns(4,5)P2 and the origin are marked.
