SecA which is an overall acidic protein was found to induce an increase in the turbidity of a solution of vesicles consisting of negatively charged phospholipids. This increase was found to be due to an aggregation of the vesicles mediated by SecA. The SecA-mediated vesicle aggregation was not found for zwitterionic 1 ,2-dioleoyl-~?~-glycero-3-phosphocholine and showed a large dependence on both temperature and ionic strength. Furthermore it was shown that ATP and to a lesser extent ADP+P, were able to reduce the SecA-mediated vesicle aggregation, while no effect could be seen for a non-hydrolysable ATP analog AMP-PNP. Usmg the steady state fluorescence anisotropy of l,h-diphenyl-1,3,5-hexatriene present m 1,2-dioleoyl-sn-glycero-3-phosphoglycerol vesicles we could show that SecA Inserts in the bllayer. Monolayer studies confirmed that SecA IS able to cause close contact between two membranes and gave a direct insight mto the different types of lipid-protein interactions involved. From our results we propose that the SecA monomer possesses two lipid-binding sites which m the functional dimer conformation are responsible for the SecA-mediated vesicle aggregation.