Great progress has been made in the elucidation of the function of proteins in membrane traffic. Less is known about the regulatory role of lipids in membrane dynamics. Studies of nerve terminals, compartments highly specialized for the recycling of synaptic vesicles, have converged with studies from other systems to reveal mechanisms in protein-lipid interactions that affect membrane shape as well as the fusion and fission of vesicles. Phosphoinositides have emerged as major regulators of the binding of cytosolic proteins to the bilayer. Phosphorylation on different positions of the inositol ring generates different isomers that are heterogeneously distributed on cell membranes and that together with membrane proteins generate a “dual keys” code for the recruitment of cytosolic proteins. This code helps controlling vectoriality of membrane transport. Powerful methods for the detection of lipids are rapidly advancing this field, thus complementing the broad range of information about biological systems that can be obtained from genomic and proteomic approaches.