Detection of submicron-sized raft-like domains in membranes by small-angle neutron scattering.
Using coarse grained models of heterogeneous vesicles we demonstrate the potential for small-angle neutron scattering (SANS) to detect and distinguish between two different categories of lateral segregation: 1) unilamellar vesicles (ULV) containing a single domain and 2) the formation of several small domains or "clusters" (approximately 10 nm in radius) on a ULV. Exploiting the unique sensitivity of neutron scattering to differences between hydrogen and deuterium, we show that the liquid ordered (lo) DPPC-rich phase can be selectively labeled using chain deuterated dipalymitoyl phosphatidylcholine (dDPPC), which greatly facilitates the use of SANS to detect membrane domains. SANS experiments are then performed in order to detect and characterize, on nanometer length scales, lateral heterogeneities, or so-called "rafts", in approximately 30 nm radius low polydispersity ULV made up of ternary mixtures of phospholipids and cholesterol. For 1:1:1 DOPC:DPPC:cholesterol (DDC) ULV we find evidence for the formation of lateral heterogeneities on cooling below 30 degrees C. These heterogeneities do not appear when DOPC is replaced by SOPC. Fits to the experimental data using coarse grained models show that, at room temperature, DDC ULV each exhibit approximately 30 domains with average radii of approximately 10 nm.
Pencer
J
J
National Research Council, Canadian Neutron Beam Centre, Chalk River, ON K0J 1J0, Canada. Jeremy.Pencer@nrc.gc.ca
Mills
T
T
Anghel
V
V
Krueger
S
S
Epand
R M
RM
Katsaras
J
J
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D055592:Biophysical Phenomena; D001703:Biophysics; D002784:Cholesterol / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry; D006859:Hydrogen / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008563:Membrane Lipids / Q000737:chemistry*; D036103:Nanotechnology; D033363:Neutron Diffraction / Q000379:methods*; D010316:Particle Size; D010743:Phospholipids / Q000737:chemistry; D012542:Scattering, Radiation; D013696:Temperature
D008051:Lipid Bilayers; D008563:Membrane Lipids; D010743:Phospholipids; D015060:1,2-Dipalmitoylphosphatidylcholine; D006859:Hydrogen; D002784:Cholesterol; D003903:Deuterium