Reference - Detail
| RRC ID | 72947 |
|---|---|
| Author | Fujii M, Ohtsubo M, Ogawa T, Kamata H, Hirata H, Yagisawa H. |
| Title | Real-time visualization of PH domain-dependent translocation of phospholipase C-delta1 in renal epithelial cells (MDCK): response to hypo-osmotic stress. |
| Journal | Biochem Biophys Res Commun |
| Abstract |
Green fluorescent protein (GFP)-tagged phospholipase C (PLC)-delta1 and its mutants were expressed in Madin-Darby canine kidney (MDCK) cells. GFP-PLC-delta1 or the GFP-tagged pleckstrin homology (PH) domain of PLC-delta1 itself was found to be predominantly localized at the plasma membrane. The DeltaPH mutant or a site-directed mutant containing a PH domain which does not bind inositol 1,4, 5-trisphosphate and cannot hydrolyze phosphatidylinositol 4, 5-bisphosphate in vitro was seen only in the cytosol. In living MDCK cells hypo-osmotic stress caused a rapid dissociation of GFP-PLC-delta1 from the plasma membrane, which coincided with phosphoinositide breakdown. A PLC inhibitor, U73122, blocked this translocation, but depletion of extracellular Ca2+ had no effect. The translocation was reversed by replacement with an iso-osmotic buffer. Our results demonstrate that the PH domain plays a critical role in the membrane targeting of PLC-delta1 and that the intracellular distribution of the enzyme is regulated by osmotic stress-driven phosphoinositide turnover. |
| Volume | 254(2) |
| Pages | 284-91 |
| Published | 1999-1-19 |
| DOI | 10.1006/bbrc.1998.9936 |
| PII | S0006-291X(98)99936-3 |
| PMID | 9918830 |
| MeSH | Animals Calcium / metabolism Catalytic Domain Cell Line Cell Membrane / enzymology* Cytosol / enzymology Dogs Enzyme Inhibitors / pharmacology Epithelial Cells Estrenes / pharmacology Green Fluorescent Proteins Hypotonic Solutions Inositol 1,4,5-Trisphosphate / metabolism Isoenzymes / chemistry* Isoenzymes / metabolism* Kidney Kinetics Luminescent Proteins / metabolism Phospholipase C delta Pyrrolidinones / pharmacology Recombinant Fusion Proteins / metabolism Substrate Specificity Time Factors Transfection Type C Phospholipases / chemistry* Type C Phospholipases / metabolism* src Homology Domains |
| IF | 2.985 |
| Resource | |
| Human and Animal Cells | MDCK(RCB0995) |