Solubilization and reconstitution of membrane energy-transducing systems ofEscherichia coli
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چکیده
منابع مشابه
Cation/proton antiport systems in Escherichia coli. Solubilization and reconstitution of delta pH-driven sodium/proton and calcium/proton antiporters.
Uptake of 22Na+ and 45Ca2+ into everted membrane vesicles from Escherichia coli was measured with imposed transmembrane pH gradients, acid interior, as driving force. Vesicles loaded with 0.5 M KCl were diluted into 0.5 M choline chloride to create a potassium gradient. Addition of nigericin to produce K+/H+ exchange resulted in formation of a pH gradient. This imposed gradient was capable of d...
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A strain of EScherichia coli was constructed containing a plasmid from the Clarke-Carbon collection that showed high levels of melibiose transport activity. Membranes from the plasmid-containing strain were extracted with octyl-beta-D-glucopyranoside and melibiose transport was reconstituted in liposomes. The proteoliposomes exhibited counterflow activity, as well as membrane potential and sodi...
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The membrane carrier for L-proline (product of the putP gene) of Escherichia coli K12 was solubilized and functionally reconstituted with E. coli phospholipid by the cholate dilution method. The counterflow activity of the reconstituted system was studied by preloading the proteoliposomes with either L-proline or the proline analogues: L-azetidine-2-carboxylate or 3,4-dehydro-L-proline. The dil...
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The SecY protein and other membrane proteins of Escherichia coli were solubilized by mixed micelles of n-octyl beta-D-glucopyranoside, phospholipids, and glycerol. Proteoliposomes formed from this extract by detergent dialysis supported energy-dependent translocation and processing of pro-OmpA. Translocation required ATP, SecY, and SecA and was stimulated by a proton-motive force. These results...
متن کاملSolubilization and reconstitution of ca pump from corn leaf plasma membrane.
The Ca(2+) transport system of corn (Zea mays) leaf plasma membrane is composed of Ca(2+) pump and Ca(2+)/H(+) antiporter driven by H(+) gradient imposed by a H(+) pump (M Kasai, S Muto [1990] J Membr Biol 114: 133-142). It is necessary for characterization of these Ca(2+) transporters to establish the procedure for their solubilization, isolation, and reconstitution into liposomes. We attempte...
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ژورنال
عنوان ژورنال: FEBS Letters
سال: 1982
ISSN: 0014-5793
DOI: 10.1016/0014-5793(82)80141-5