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At least 541 records · Page 30Linked to original sources
PubMed1984

Simplex optimization in biochemistry: application of the method in two-phase partition.

Partition in aqueous two-phase systems should have great potentialities for studies of biological material. However, the general use of the method has probably been hindered by the difficulties in finding the composition of two-phase systems with desired qualities. Experimental design has proved to be of immense value for optimization in many different areas and should also be useful in selecting the best possible two-phase system. Therefore the simplex method has been applied to the search for the composition of an aqueous two-phase system in which the partition behavior of filamentous actin and human spectrin differ as much as possible.

Actins↗
PubMed1976

Isolation of photosystem II enriched membrane vesicles from spinach chloroplasts by phase partition.

Partition in an aqueous Dextran-polyethylene glycol two-phase system has been used for the separation of chloroplast membrane vesicles obtained by press treatment of a grana-enriched fraction after unstacking in a low salt buffer. The fractions obtained were analysed with respect to chlorophyll, photochemical activities and ultrastructural charasteristics. The results reveal that the material partitioning to the Dextran-rich bottom phase consisted of large membrane vesicles possessing mainly Photosystem II properties with very low contribution from Photosystem I. Measurements of the H2O to phenyl-p-benzoquinone and ascorbate-Cl2Ind to NADP+ electron transport rates indicate a ratio of around six between Photosystem II and I. The total fractionation procedure could be completed within 2-3 h with high recovery of both the Photosystem II water-splitting activity and the Photosystem I reduction of NADP+. These data demonstrate that press treatment of low-salt destabilized grana membranes yields a population of highly Photosystem-II enriched membrane vesicles which can be discriminated by the phase system. We suggest that such membrane vesicles originate from large regions in the native grana membrane which contain virtually only Photosystem II.

Cell Fractionation↗
PubMed1978

Synaptic membranes from Torpedo marmorata electric organ. 1. Separation and analysis of nicotinic acetylcholine receptor- and acetylcholinesterase-containing membrane vesicles using aqueous two-phase systems.

Two-phase systems consisting of water, dextran and poly(ethylene glycol) have been used for partition of membranes obtained from Torpedo marmorata electric organ. The partition behaviour of the membranes could be adjusted by using a polymer with covalently-bound charged groups. By using this method, the membranes were divided into several fractions which were analyzed for nicotinic acetylcholine receptor and acetylcholinesterase content. It was found that nicotinic receptor-enriched membranes were separated from those containing esterase in a single partition step. Receptor-enriched membranes obtained by gradient centrifugation could be further separated into two receptor fractions by the two-phase technique. The results also reveal at least two types of acetylcholinesterase-rich membranes.

Acetylcholinesterase↗
PubMed1979

Effect of membrane cholesterol enrichment or depletion on the partition behavior of human erythrocytes in dextran-poly(ethylene glycol) aqueous phases.

It has previously been shown that by appropriate manipulation of polymer concentrations and ionic composition and concentration one can select whether charge-associated or lipid-related membrane surface properties are reflected by cell partition in dextran-poly(ethylene glycol) aqueous two-phase systems (Walter, H. (1977) in Methods of Cell Separation ((Catsimpoolas, N., ed.), Vol. 1, pp. 307-354, Plenum Press, New York). In the current experiments we have studied that partition behavior of human erythrocytes and found that not only lipid-related but also charge-associated membrane properties are altered as a consequence of cholesterol-enrichment or -depletion. Results further indicate that, just as cell partition in charged phase systems reflects membrane charge-associated properties not readily measured by means other than partition (Brooks, D.E., Seaman, G.V.F. and Walter, H. (1971) Nat. New Biol. 234, 61--62; Walter, H., Tung, R., Jackson, L.J. and Seaman, G.V.F. (1972) Biochem. Biophys. Res. Commun. 48, 565--571), cell partition in uncharged phases reflects membrane lipid-related properties also not readily measured by other means.

Cell Separation↗