Details

Autor(en) / Beteiligte

• Adjobo-Hermans, Merel J.W
• Goedhart, Joachim
• Gadella, Theodorus W.J. Jr

Titel

Regulation of PLCβ1a membrane anchoring by its substrate phosphatidylinositol (4,5)-bisphosphate

Ist Teil von

• Journal of cell science, 2008-11, Vol.121 (22), p.3770-3777

Ort / Verlag

The Company of Biologists Limited

Erscheinungsjahr

2008

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• Basic knowledge as to the subcellular location and dynamics of PLCβ isozymes is scant. Here, we report on the subcellular location of GFP-PLCβ1a and the use of total internal reflection fluorescence (TIRF) microscopy to examine the dynamics of GFP-PLCβ1a at the plasma membrane upon stimulation of Gq-coupled receptors. Using this technique, we observed PLCβ1a dissociation from the plasma membrane upon addition of agonist. An increase in intracellular calcium and a decrease in PtdIns(4,5)P₂ both coincided with a translocation of PLCβ1a from the plasma membrane into the cytosol. In order to differentiate between calcium and PtdIns(4,5)P₂, rapamycin-induced heterodimerization of FRB and FKBP12 fused to 5-phosphatase IV was used to instantaneously convert PtdIns(4,5)P₂ into PtdIns(4)P. Addition of rapamycin caused PLCβ1a to dissociate from the plasma membrane, indicating that removal of PtdIns(4,5)P₂ is sufficient to cause translocation of PLCβ1a from the plasma membrane. In conclusion, PLCβ1a localization is regulated by its own substrate.