Reconstitution of GTP‐γ‐S‐dependent phospholipase D activity with ARF, RhoA, and a soluble 36‐kDa protein

[摘要]

For activation of kidney membrane phospholipase D (PLD), cytosol is absolutely needed in addition to GTP-γ-S. The active component of cytosol consists of three protein factors: ADP-ribosylation factor, RhoA, and a soluble 36-kDa protein. Any combination of these two factors synergistically activates PLD to some extent, but the presence of the three factors causes full activation. The 36-kDa protein is stable at 60°C but inactivated at 80°C for 10 min. Tissue distribution of the 36-kDa protein roughly coincides with that of PLD, suggesting physiological relevance of the protein in the regulation of PLD.

[学科分类] 生物化学/生物物理

[关键词] Phospholipase D;ADP-ribosylation factor;RhoA;G-protein;PLD;phospholipase D;PtdCho;phosphatidylcholine;G-protein;GTP-binding regulatory protein;ARF;ADP-ribosylation factor;GTP-γ-S;guanosine 5′-O-(3-thiotriphosphate);PtdEtn;phosphatidylethanolamine;PtdEtOH;phosphatidylethanol;PKC;protein kinase C