title: Influence of liposome composition and membrane binding on protein kinase activity of PI3Kγ.
abstract: Phosphoinositide 3-kinase γ (PI3Kγ) has been implicated in a variety of cellular signaling processes. It is a multifunctional enzyme with lipid and protein kinase activity that also acts as a scaffold protein. Although it is well known that membrane recruitment is essential for the phosphorylation of phosphoinositides, the cellular localization of PI3Kγ as a protein kinase remains unclear. It has merely been described that PI3Kγ protein kinase activity leading to MAPK activation seems to be restricted to a cytosolic localization. Here, we demonstrate that a hybrid-PI3Kγ having protein kinase, but not lipid kinase activity shows a similar cellular distribution with a high membrane association and comparable liposome binding behavior to wild-type PI3Kγ. Binding of PI3Kγ to liposomes mimicking the natural plasma membrane slightly stimulates autophosphorylation of PI3Kγ. However, liposomes containing an unphysiologically high amount of PI inhibit autophosphorylation of PI3Kγ. Finally, PI3Kγ bound to membrane fragments does not show autophosphorylation which is possibly due to protein-protein-interactions at the plasma membrane. This indicates that not only MAPK activation, but PI3Kγ protein kinase activity in general is localized in the cytosol.
authors.affiliation: Institute of Biochemistry II, Medical Faculty, Friedrich-Schiller-University Jena, Nonnenplan 4, Jena 07740, Germany. christinadolle@aol.com
authors.forename: Christina
authors.initials: C
authors.lastname: Dolle
authors.forename: Martin
authors.initials: M
authors.lastname: Westermann
authors.forename: Margret
authors.initials: M
authors.lastname: Schilli-Westermann
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kirsch
mesh_terms: D002462:Cell Membrane / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology*; D057809:HEK293 Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism
chemical_list: D008081:Liposomes; D019869:Phosphatidylinositol 3-Kinases; D010743:Phospholipids; D058570:TOR Serine-Threonine Kinases; C546842:MTOR protein, human; C525865:PIK3C2G protein, human