Neuronal exocytosis is mediated by Ca21-triggered rearrangementsbetween proteins and lipids that result in the opening anddilation of fusion pores. Synaptotagmin I (syt I) is a Ca21-sensingprotein proposed to regulate fusion pore dynamics via Ca21-promoted binding of its cytoplasmic domain (C2A-C2B) to effectormolecules, including anionic phospholipids and other copies of syt.Functional studies indicate that Ca21-triggered oligomerization ofsyt is a critical step in excitation–secretion coupling; however, thisactivity has recently been called into question. Here, we show thatCa21 does not drive the oligomerization of C2A-C2B in solution.However, analysis of Ca21zC2A-C2B bound to lipid monolayers,using electron microscopy, revealed the formation of ring-likeheptameric oligomers that are '11 nm long and '11 nm indiameter. In some cases, C2A-C2B also assembled into long filaments.Oligomerization, but not membrane binding, was disruptedby neutralization of two lysine residues (K326,327) within the C2Bdomain of syt. These data indicate that Ca21 first drives C2AC2Bzmembraneinteractions, resulting in conformational changesthat trigger a subsequent C2B-mediated oligomerization step.Ca21-mediated rearrangements between syt subunits may regulatethe opening or dilation kinetics of fusion pores or may play arole in endocytosis after fusion.

SecA, an essential component of the general protein secretion pathwayof bacteria, is present in Escherichia coli as soluble andmembrane-integral forms. Hereweshow by electron microscopy thatSecA assumes two characteristic forms in the presence of phospholipidmonolayers: dumbbell-shaped elongated structures and ring-likepore structures. The ring-like pore structures with diameters of 8 nmand holes of 2nm are found only in the presence of anionic phospholipids.These ring-like pore structures with larger 3- to 6-nm holes(without staining) were also observed by atomic force microscopicexamination. They do not form in solution or in the presence ofuncharged phosphatidylcholine. These ring-like phospholipidinducedpore-structures may form the core of bacterial proteinconductingchannels through bacterial membranes.

β-Amyloid peptide (Aβ), a normal constituent of neuronal and non-neuronal cells, has been proven to be the major component of extracellular plaque of Alzheimer's disease. Interactions between Aβand neuronal membranes have been postulated to play an important role in the neuropathology of Alzheimer’s disease. Here we show that Aβ is able to insert into lipid bilayer. The membrane insertion ability of Aβ is critically controlled by the ratio of cholesterol to phospholipids. In a low concentration of cholesterol Aβ prefers to stay in membrane surface region mainly in aβ-sheet structure. In contrast, as the ratio of cholesterol to phospholipids rises above 30mol%, Aβ can insert spontaneously into lipid bilayer by its C terminus. During membrane insertion Aβ generates about 60%β-helix and removes almost all β-sheet structure. Fibril formation experiments show that such membrane insertion can reduce fibril formation. Our findings reveal a possible pathway by which Aβ prevents itself from aggregation and fibril formation by membrane insertion.

Many proteins exhibit both a water-soluble and a membrane-bound state. The proteins in the membrane-bound state obtain a distinct structure from that in the bulk, which exists in many important biological processes. In the present paper we would stress that the variation of the physical chemistry properties of the microenvironment adjacent to the membranesurface region play an important role in the process of the membrane-induced conformational changes of the proteins.

The interaction of rabbit C-reactive protein (rCRP) with a supported monolayer containing a phosphorylcholine moiety was studied. Three types of phospholipids were synthesized, each containing a insertion spacer of eight, six, or three atoms between the phosphorylcholine group and hydrophobic tail. By varying the length of the insertion spacer, we can vary the extension of the phosphorylcholine group from the membrane surface. By varying the monolayer composition, we can control the lateral distance between the exposed phosphorylcholine groups. Using the surface plasmon resonance technique (SPR), we demonstrated that the calcium-dependent binding of rCRP to the model membrane is governed not only by the ability of the ligand to access the binding pocket fully (spacer length), but also by lateral hindrance within the two-dimensional plane of the membrane. The value of the apparent binding constant was estimated by theoretical analysis, which is obviously dependent on the composition of the lipid mixture, and a maximum of (9.9 6 1.5) 3 106 M21 was obtained.