In this section
@article{Mulet:2009:10.1007/978-1-60327-115-8_8,
author = {Mulet, X and Rosivatz, E and Ho, KK and Gauthe, BL and Ces, O and Templer, RH and Woscholski, R},
doi = {10.1007/978-1-60327-115-8_8},
journal = {Methods Mol.Biol.},
pages = {135--144},
title = {Spatial localization of PtdInsP2 in phase-separated giant unilamellar vesicles with a fluorescent PLC-delta 1 PH domain},
url = {http://dx.doi.org/10.1007/978-1-60327-115-8_8},
volume = {462},
year = {2009}
}
TY - JOUR
AB - This chapter describes a method for the preparation of giant unilamellar vesicles containing phosphatidylinositol 4,5-bisphosphate that are larger than 20 microm in size. The phospholipids composition of the vesicular membrane is such that fluid lamellar and liquid-ordered or gel phases are formed and separate within the confines of one vesicle. It outlines the preparation of a protein fluorescent label, pleckstrin homology domain from phospholipase C-delta 1, that binds specifically to phosphatidylinositol 4,5-bisphosphate. Using fluorescence microscopy, the presence and spatial position of this phosphorylated phosphatidylinositol lipid on the lipid membrane have been located with the pleckstrin homology domain. We show that phosphatidylinositol 4,5-bisphosphate and the phospholipase C-delta 1 pleckstrin homology domain are located to the fluid phase of the vesicle membrane. This approach can therefore show how membrane physical properties can affect enzyme binding to phosphatidylinositol 4,5-bisphosphate and thus further the understanding of important membrane processes such as endocytosis
AU - Mulet,X
AU - Rosivatz,E
AU - Ho,KK
AU - Gauthe,BL
AU - Ces,O
AU - Templer,RH
AU - Woscholski,R
DO - 10.1007/978-1-60327-115-8_8
EP - 144
PY - 2009///
SP - 135
TI - Spatial localization of PtdInsP2 in phase-separated giant unilamellar vesicles with a fluorescent PLC-delta 1 PH domain
T2 - Methods Mol.Biol.
UR - http://dx.doi.org/10.1007/978-1-60327-115-8_8
UR - pm:19160665
VL - 462
ER -
