Isolation of plasma membrane

Keenan, T.W., Valivullah, H.M., Dunlevy, J.T. 1989. Isolation of plasma membranes from mammary gland by two phase polymer partitioning. Anal. Biochem. 177, 194-198. 

WaUach DF, Schmidt-Ullrich R. Isolation of plasma membrane vesicles from animal cells. Methods Cell Biol. 1977 15 235-276. Brown DA. Lipid rafts, detergent-resistant membranes, and raft targeting signals. Physiology 2006 21 430-439. 

Mersel, M., et al. (1987). Isolation of Plasma Membranes from Neurons Grown in Primary Culture, Anal. Biochem. 166 246-252. 

Scott, L., Schell, M. J., and Hubbard, A. L. (1993). Isolation of plasma membrane sheets and plasma membrane domains from rat liver. Methods Mol. Biol 19, 59-69. 

Blowers, D.P., Hetherington, A. and Trewavas, A. 1985. Isolation of plasma membrane bound calcium/calmodulin regulated protein kinase from pea using western blotting. Planta 166, 208-15. 

Isolation of plasma membranes The plasma membrane-enriched fractions from the respective treatments were collected on sucrose gradients as described before (Sancholle 1984 Fonvieille 1985). The sucrose concentrations from the bottom of the centrifuge tube were 50% (4.8 ml),... 

Wang, G.-D. Stucfy of isolation of plasma membrane, characterization of plasma membrane IT -ATPase and effect of in vitro peroxidation on plasma membrane from bell pepper fruit tissue (Capsicum annum L.) fruit tissue. M.Sc. Thesis, University of Georgia, Athens, 1992. 

Auderset, G., Sandelius, A. S., Penel, C., Brightman, A., Greppin, H., and MorrI, D. J., Isolation of plasma membrane and tonoplast fractions from spinach leaves by preparative free-flow electrophoresis and effect of photoinduction. Physiol. Plant, (in press). 

Boone, C. W., Ford, L. E., Bond, H. E., Stuart, D. C., and Lorenz, D., 1969, Isolation of plasma membrane fragments from HeLa cells, /. Cell. Biol. 41 378. 

Kidwai, A. M., Radcliffe, M. A., Duchon, G., and Daniel, E. E., 1971, Isolation of plasma membrane from cardiac muscle, Biochem. Biophys. Res. Commun. 45 901. 

M. J. Holden, D. G. Luster, R. L. Chaney, T. J. Buckhout. and C. Robinson, Fc -chelate reductase activity of plasma membranes isolated from tomato (Lyco-persicon escidentiim Mill.) roots. Comparison of enzymes from Fe-deficient and Fe-sufficient roots. Plant Physiol. 97 531 (1991). 

Z. Varanini, R. Pinton, M. G. De Biasi, S. Astolfi, and A. Maggioni, Low molecular weight humic substances stimulate H -ATPase activity of plasma membrane vesicles isolated from oat (Avena sativa L.) roots. Plant Soil I53 6 (1993). 

Possible explanations for a blood flow-limited uptake in kidney include the existence of specific uptake mechanisms, such as receptor-mediated endocytosis and carrier-mediated transport. Since the former mechanism is initiated by binding of the ligand to the cell-surface receptor, the specific binding of alkylglycoside compounds to isolated tubular plasma membranes was examined [23,24]. 

TABLE 9.1. Effect of Different Humic Substances Fractions (HS) on Active Proton Extrusion from Intact Roots and on H+-ATPase Activty of Plasma Membrane (PM) Vesicles Isolated from Different Plant Roots... 

Matthyssee and Phillips (20) isolated two nuclear proteins, from tobacco cells, that bound specifically to 2,4-D. Receptor proteins for auxins, kinetins, and GA have been found (21). Sub-cellular fractions from bean leaves were recently shown to bind abscisic acid (22). Preliminary experiments (22) indicated that maximum ABA binding activity coincides with the activities of membrane-bound Mg -dependent, K+-stimulated ATPase and glucan synthetase. Table I of Biswas and Roy (21) lists hormone receptor proteins reported in plant tissue. For a protein to qualify as a receptor molecule, it should have a high stereo-specific binding capacity (Kd 10 6 to 10 SM) for its particular hormone. In com coleoptiles, both IAA and NAA are equally effective in inducing cell elongations fractions of plasma membrane and endoplasmic reticular membrane contain receptor proteins with Kd values of 10 M to 10 M for auxins (5, 18). When one considers procedural... 

A hormone receptor was isolated from plasma membranes of liver cells, and its molecular weight was found to be 120,000. It was incubated at a concentration of 2 mg/mL with various concentrations of the hormone, and this was followed by measurements at equilibrium of free and bound A (in mol/L) at 37 and 4°C. The data are presented here. Answer Questions 16.8 and 16.9 on the basis of this information. 

The preparation of plasma membrane vesicles from liver canalicular membrane is highly enriched with the canalicular (apical) isoform MRP2 (Buchler et al. 1996). Methods for the isolation of hepatocyte canalicular membranes from liver tissue have been described in detail (Bohme et al. 1994 and Boyer and Meier et al. 1990). The percentage of inside-out-oriented vesicles in these preparations amounts to 32%. Alternatively, transfected I ILK and MDCK cells are often used to study ATP-dependent transport into inside-out vesicles (Cui et al. 1999 Leier et al. 2000). 

Figure 13.5 Schematic representation of the fractionation and analysis of plasma membrane proteins isolated using the colloidal silica method. Figure provided by A. Rahbar.

Melin, P.M., Sommarin, M., Sandelius, A.S., and Jergil, B., 1987, Identification of Ca2+-stimulated polyphosphoinositide phospholipase C in isolated plant plasma membranes. FEBS Lett. 223 87-91. 

Bontemps, F., van den Berghe, G. Metabolism of exogenous S-adeno-sylmethionine in isolated rat hepatocyte suspensions methylation of plasma-membrane phospholipids without intracellular uptake. Bio-... 

C, R. Ross and P D. Holohan, liansport of organic atuons and cations in isolated renal plasma membranes, Ann. Rev. Pharmacol. Toxicol, 23 65-85 (1983). 

For shotgnn-LC-MS identification, selective prefractionation by means of AfC is a powerful tool. Biotinylation of cell-surface proteins and subsequent streptavidin-AfC enabled a 1600-fold and 400-fold emichment of plasma membrane proteins over proteins from mitochondria and endoplasmic reticulum, respectively [78-79]. Because of the selective isolation procedure, only 3% of the 918 unique proteins identified were related to mitochondria and endoplasmic reticulum proteins, and 25% were either plasma membrane proteins or membrane-associated proteins. 

Carafoli, E. (1984). Calmodulin-sensitive calcium-pumping ATPase of plasma membranes Isolation, reconstitution and regulation. Fed. Proc. 43, 3005-3010. 

Staining with PI is optional (e.g., Fig. 6). It allows to distinguish the cells that have integrity of plasma membrane compromised to the extent that they cannot exclude PI (necrotic and late apoptotic cells, cells with mechanically damaged membranes, isolated cell nuclei). 

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