title: Peripheral protein organization and its influence on lipid diffusion in biomimetic membranes.
abstract: Protein organization on biomembranes and their dynamics are essential for cellular function. It is not clear, however, how protein binding may influence the assembly of underlying lipids or how the membrane structure leads to functional protein organization. Toward this goal, we investigated the effects of annexin a5 binding to biomimetic membranes using fluorescence imaging and correlation spectroscopy. Annexin a5 (anx a5), a peripheral intracellular protein that plays a membrane remodeling role in addition to other functions, binds specifically and tightly to anionic (e.g., phosphatidylserine)-containing membranes in the presence of calcium ion. Our fluorescence microscopy reveals that annexin likely forms assemblies, along with a more dispersed population, upon binding to anionic biomembranes in the presence of calcium ion, which is reflected in its two-component Brownian motion. To investigate the effects of annexin binding on the underlying lipids, we used specific acyl chain labeled phospholipid analogues, NBD-phosphatidylcholine (NBD-PC) and NBD-phosphatidylserine (NBD-PS). We find that both NBD-labeled lipids cluster under anx a5 assemblies, as compared with when they are found under the dispersed annexin population, and NBD-PS exhibits two-component lateral diffusion under the annexin assemblies. In contrast, NBD-PC diffusion is slower by an order of magnitude under the annexin assemblies in contrast to its diffusion when not localized under anx a5 assemblies. Our results indicate that, upon binding to membranes, the peripheral protein annexin organizes the underlying lipids into domains, which may have functional implications in vivo.
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Kanika
authors.initials: K
authors.lastname: Vats
authors.forename: Kristofer
authors.initials: K
authors.lastname: Knutson
authors.forename: Anne
authors.initials: A
authors.lastname: Hinderliter
authors.forename: Erin D
authors.initials: ED
authors.lastname: Sheets
mesh_terms: D017304:Annexin A5 / Q000737:chemistry / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D004058:Diffusion; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence
chemical_list: D017304:Annexin A5; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010718:Phosphatidylserines; C050294:1-palmitoyl-2-oleoylglycero-3-phosphoserine; D002118:Calcium