TY - JOUR
T1 - Use of the GRP1 PH domain as a tool to measure the relative levels of PtdIns(3,4,5)P3 through a protein-lipid overlay approach
AU - Guillou, Hervé
AU - Lécureuil, Charlotte
AU - Anderson, Karen E.
AU - Suire, Sabine
AU - Ferguson, G. John
AU - Ellson, Chris D.
AU - Gray, Alexander
AU - Divecha, Nullin
AU - Hawkins, Phillip T.
AU - Stephens, Len R.
PY - 2007/3
Y1 - 2007/3
N2 - We describe a novel approach to the relative quantification of phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P3] and its application to measure, in neutrophils, the activation of phosphoinositide 3-kinase (PI3K). This protein-lipid overlay-based assay allowed us to confirm and extend the observations, first, that N-formyl-methionylleucyl-phenylalanine (fMLP) stimulation of primed human neutrophils leads to a transient and biphasic increase in PtdIns(3,4,5)P3 levels and, second, that the ability of fMLP to stimulate PtdIns(3,4,5)P3 accumulation in neutrophils isolated from mice carrying a Ras-insensitive ('DASAA') knock-in of PI3Kγ (p110γDASAA/DASAA) is substantially dependent on the Ras binding domain of PI3Kγ.
AB - We describe a novel approach to the relative quantification of phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P3] and its application to measure, in neutrophils, the activation of phosphoinositide 3-kinase (PI3K). This protein-lipid overlay-based assay allowed us to confirm and extend the observations, first, that N-formyl-methionylleucyl-phenylalanine (fMLP) stimulation of primed human neutrophils leads to a transient and biphasic increase in PtdIns(3,4,5)P3 levels and, second, that the ability of fMLP to stimulate PtdIns(3,4,5)P3 accumulation in neutrophils isolated from mice carrying a Ras-insensitive ('DASAA') knock-in of PI3Kγ (p110γDASAA/DASAA) is substantially dependent on the Ras binding domain of PI3Kγ.
KW - General receptor for phosphoinositides-1
KW - Neomycin
KW - Neutrophil
KW - p110γ
KW - Phosphatidylinositol (3,4,5)-trisphosphate
KW - Phosphoinositide 3-kinase
KW - Protein-lipid overlay
KW - Ras
U2 - 10.1194/jlr.D600038-JLR200
DO - 10.1194/jlr.D600038-JLR200
M3 - Article
C2 - 17130283
AN - SCOPUS:33947174209
VL - 48
SP - 726
EP - 732
JO - Journal of Lipid Research
JF - Journal of Lipid Research
SN - 0022-2275
IS - 3
ER -