Membranes microscopy

Micellar structure has been a subject of much discussion [104]. Early proposals for spherical [159] and lamellar [160] micelles may both have merit. A schematic of a spherical micelle and a unilamellar vesicle is shown in Fig. Xni-11. In addition to the most common spherical micelles, scattering and microscopy experiments have shown the existence of rodlike [161, 162], disklike [163], threadlike [132] and even quadmple-helix [164] structures. Lattice models (see Fig. XIII-12) by Leermakers and Scheutjens have confirmed and characterized the properties of spherical and membrane like micelles [165]. Similar analyses exist for micelles formed by diblock copolymers in a selective solvent [166]. Other shapes proposed include ellipsoidal [167] and a sphere-to-cylinder transition [168]. Fluorescence depolarization and NMR studies both point to a rather fluid micellar core consistent with the disorder implied by Fig. Xm-12. 

Phospholipid molecules form bilayer films or membranes about 5 nm in thickness as illustrated in Fig. XV-10. Vesicles or liposomes are closed bilayer shells in the 100-1000-nm size range formed on sonication of bilayer forming amphiphiles. Vesicles find use as controlled release and delivery vehicles in cosmetic lotions, agrochemicals, and, potentially, drugs. The advances in cryoelec-tron microscopy (see Section VIII-2A) in recent years have aided their characterization [70-72]. Additional light and x-ray scattering measurements reveal bilayer thickness and phase transitions [70, 71]. Differential thermal analysis... 

The interest in vesicles as models for cell biomembranes has led to much work on the interactions within and between lipid layers. The primary contributions to vesicle stability and curvature include those familiar to us already, the electrostatic interactions between charged head groups (Chapter V) and the van der Waals interaction between layers (Chapter VI). An additional force due to thermal fluctuations in membranes produces a steric repulsion between membranes known as the Helfrich or undulation interaction. This force has been quantified by Sackmann and co-workers using reflection interference contrast microscopy to monitor vesicles weakly adhering to a solid substrate [78]. Membrane fluctuation forces may influence the interactions between proteins embedded in them [79]. Finally, in balance with these forces, bending elasticity helps determine shape transitions [80], interactions between inclusions [81], aggregation of membrane junctions [82], and unbinding of pinched membranes [83]. Specific interactions between membrane embedded receptors add an additional complication to biomembrane behavior. These have been stud-... 

Knoll G and Plattner H 1989 Ultrastructural analysis of biological membrane fusion and a tentative correlation with biochemical and biophysical aspects Electron Microscopy of Subcellular Dynamics ed H Plattner (London CRC) pp 95-117... 

Brandow S L, Turner D C, Ratna B R and Gaber B P 1993 Modification of supported lipid membranes by atomic force microscopy Biophys. J. 64 898... 

Two-photon excited fluorescence detection at the single-molecule level has been demonstrated for cliromophores in cryogenic solids [60], room-temperature surfaces [61], membranes [62] and liquids [63, 64 and 65]. Altliough multiphoton excited fluorescence has been embraced witli great entluisiasm as a teclmique for botli ordinary confocal microscopy and single-molecule detection, it is not a panacea in particular, photochemical degradation in multiphoton excitation may be more severe tlian witli ordinary linear excitation, probably due to absorjDtion of more tlian tire desired number of photons from tire intense laser pulse (e.g. triplet excited state absorjDtion) [61],... 

Hollars C W and Dunn R C 2000 Probing single molecule orientations in model lipid membranes with near-field scanning optical microscopy J. Phys. Chem 112 7822-30... 

The spatial arrangement of atoms in two-dimensional protein arrays can be detennined using high-resolution transmission electron microscopy [20]. The measurements have to be carried out in high vacuum, but since tire metliod is used above all for investigating membrane proteins, it may be supposed tliat tire presence of tire lipid bilayer ensures tliat tire protein remains essentially in its native configuration. 

Two-dimensional crystals of membrane proteins can be studied by electron microscopy... 

This electron microscopy reconstruction has since been extended to high resolution (3 A) where the connections between the helices and the bound retinal molecule are visible together with the seven helices (Figure 12.3c). The helices are tilted by about 20° with respect to the plane of the membrane. This is the first example of a high-resolution three-dimensional protein structure determination using electron microscopy. The structure has subsequently been confirmed by x-ray crystallographic studies to 2 A resolution. 

The three-dimensional structure of the bacterial membrane protein, bac-teriorhodopsin, was the first to be obtained from electron microscopy of two-dimensional crystals. This method is now being successfully applied to several other membrane-bound proteins. 

Henderson, R., Unwin, RN.T. Three-dimensional model of purple membrane obtained by electron microscopy. Nature 257 28-32, 1975. 

Alphaviruses, such as Sindbis virus and Semliki Forest virus, are a group of mosquito-borne, enveloped RNA viruses that can cause encephalitis, fever, arthritis and rashes in mammals. These viruses have two protein shells—an outer glycoprotein layer and an inner core— which are separated by a lipid bilayer, a membrane. Studies by cryoelectron microscopy have shown that... 

Microscopy, Membrane filter good for microscopy gravimetric or identification of particles and counting chemical J where required. 

Ultrafiltration utilizes membrane filters with small pore sizes ranging from O.OlS t to in order to collect small particles, to separate small particle sizes, or to obtain particle-free solutions for a variety of applications. Membrane filters are characterized by a smallness and uniformity of pore size difficult to achieve with cellulosic filters. They are further characterized by thinness, strength, flexibility, low absorption and adsorption, and a flat surface texture. These properties are useful for a variety of analytical procedures. In the analytical laboratory, ultrafiltration is especially useful for gravimetric analysis, optical microscopy, and X-ray fluorescence studies. 

Membrane proteins in many cases are randomly distributed through the plane of the membrane. This was one of the corollaries of the fluid mosaic model of Singer and Nicholson and has been experimentally verified using electron microscopy. Electron micrographs show that integral membrane proteins are often randomly distributed in the membrane, with no apparent long-range order. 

Ryanodine Receptor. Figure 1 Three-dimensional architecture of the RyR1 by cryo-electron microscopy, (a), top view (from the T-tubule) (b), bottom view (from the SR lumen) (c), side view (parallel to the SR membrane). The binding sites of FKBP12, apo-CaM and Ca -CaM are indicated in the side view. Courtesy of Dr. M. Samso (modified from Samso etal. (2005) Nat Struct Mol Biol 12 539-544). 

Figure 7. Video-enhanced DIC microscopy of rat liver Golgi apparatus membrane networks moving along microtubules using Xenopus egg microtubule motors (Allan and Vale, 1994). Top panel membrane extension with a bulbous terminus (arrow) attached to a microtubule (arrow heads). Middle panel same field two seconds later. The membrane has advanced about 3 pm along the microtubule (arrow). Bottom panel membrane has now advanced further along the microtubule (arrow). Bar = 2 pm.

Affinity chromatography using factor XII as ligand leads to purification of u-PAR rather selectively, with only trace quantities of cytokeratin 1 or gClqR present [K. Joseph and A. Kaplan, unpubl. observations]. It is of interest that none of these three proteins possesses a transmembrane domain but u-PAR has a phos-phatidylinositol linkage within the cell membrane. Nevertheless, each of them has been isolated from purified cell membranes and they have been demonstrated to exist within the cell membrane by immunoelectron microscopy [41] presumably... 

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