Recruitment of AP-1 clathrin adaptors to liposomal membranes

摘要: Protein and membrane traffic between organelles within the endocytic exocytic pathway is mediated most prominently by coated vesicles. These vesicles are formed assembly of cytosolic coat proteins onto donor membrane, which deform it into a bud so that can pinch off. Clathrin with its associated adaptors, COPI COPII three major coats. Various in vitro studies allowed insight mechanism formation. vesicle budding from chemically defined liposomes has been reconstituted vitro, using pure compounds. Further, demonstrated cargo sorted these The clathrin-coated formation appears to be more complicated. AP-1 clathrin adaptor involved at transGolgi network endosomes. This work presents an assay where recruited peptidoliposomes, presenting covalently linked peptides corresponding sorting signals. In this system, recruitment depends on myristyolated ADP-ribosylation factor(ARF1), GTP or GMP-PNP, tyrosine signals small amount phosphoinositides, phosphatidyl inositol 4,5bisphosphate. such minimal system as highmolecular weight complex indicating precoat absence clathrin. hydrolysis, induced ARF GTPase-activating protein(ARFGAP1), disassembled complex. able enhance GAP activity ARFGAP1 myristoylated ARF1, suggesting regulatory function hydrolysis early steps recruitment. provides insights might also used investigate other