title: Lipid segregation and membrane budding induced by the peripheral membrane binding protein annexin A2.
abstract: The formation of dynamic membrane microdomains is an important phenomenon in many signal transduction and membrane trafficking events. It is driven by intrinsic properties of membrane lipids and integral as well as membrane-associated proteins. Here we analyzed the ability of one peripherally associated membrane protein, annexin A2 (AnxA2), to induce the formation of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2)-rich domains in giant unilamellar vesicles (GUVs) of complex lipid composition. AnxA2 is a cytosolic protein that can bind PI(4,5)P2 and other acidic phospholipids in a Ca(2+)-dependent manner and that has been implicated in cellular membrane dynamics in endocytosis and exocytosis. We show that AnxA2 binding to GUVs induces lipid phase separation and the recruitment of PI(4,5)P2, cholesterol and glycosphingolipids into larger clusters. This property is observed for the full-length monomeric protein, a mutant derivative comprising the C-terminal protein core domain and for AnxA2 residing in a heterotetrameric complex with its intracellular binding partner S100A10. All AnxA2 derivatives inducing PI(4,5)P2 clustering are also capable of forming interconnections between PI(4,5)P2-rich microdomains of adjacent GUVs. Furthermore, they can induce membrane indentations rich in PI(4,5)P2 and inward budding of these membrane domains into the lumen of GUVs. This inward vesiculation is specific for AnxA2 and not shared with other PI(4,5)P2-binding proteins such as the pleckstrin homology (PH) domain of phospholipase Cδ1. Together our results indicate that annexins such as AnxA2 can efficiently induce membrane deformations after lipid segregation, a mechanism possibly underlying annexin functions in membrane trafficking.
authors.affiliation: Institute of Biochemistry, University of Muenster, Wilhelm-Klemm-Strasse, D-48149 Muenster, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Drücker
authors.forename: Milena
authors.initials: M
authors.lastname: Pejic
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Galla
authors.forename: Volker
authors.initials: V
authors.lastname: Gerke
mesh_terms: D017306:Annexin A2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008567:Membranes, Artificial; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D054803:Phospholipase C delta / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009418:S100 Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000099248:S100 Calcium Binding Protein A10
chemical_list: D017306:Annexin A2; D008567:Membranes, Artificial; D019269:Phosphatidylinositol 4,5-Diphosphate; D054803:Phospholipase C delta; D009418:S100 Proteins; D000099248:S100 Calcium Binding Protein A10; C472822:ANXA2 protein, human; C518024:PLCD1 protein, human
keywords: Annexin; Calcium; Giant Unilamellar Vesicles; Lipid Raft; Membrane Budding; Membrane Curvature; Membrane Structure; Microdomain; Phosphatidylinositol 4,5-Bisphosphate; Phospholipid Vesicle