Biomembranes Part Q: ATP-Driven Pumps and Related Transport: by Sidney Fleischer, Becca Fleischer

By Sidney Fleischer, Becca Fleischer

The shipping volumes of the Biomembranes sequence have been initiated with Volumes a hundred twenty five and 126 of tools in Enzymology. those volumes coated delivery in micro organism, Mitochondria, and Chloroplasts. Volumes 156 and 157 hide ATP-Driven Pumps and similar Transport.
The subject of organic membrane delivery is a really well timed one simply because a powerful conceptual foundation for its knowing now exists

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Additional resources for Biomembranes Part Q: ATP-Driven Pumps and Related Transport: Calcium, Proton, and Potassium Pumps

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Res. 86, 277 (1984). i0 T. Steck and J. Kant, this series, Vol. 31, p. 172. 11 M. Michalak, K. Famulski, and E. Carafoli, J. Biol. Chem. 259, 15540 (1984). 32 Ca 2+ FLUXES AND REGULATION [3] FIG. 2. Electron micrograph of sarcolemmal vesicles. 2. 5 ttm in diameter. Magnification: 30,000×. Assay of Marker Enzymes The m o s t c o m m o n l y used p l a s m a m e m b r a n e m a r k e r is the N a + , K +A T P a s e , which can also be used to estimate the sidedness of the vesicles. Acetylcholinesterase copurifies with the N a + , K ÷ - A T P a s e , and this enz y m e can also be used to estimate the sidedness characteristics o f the vesicles.

Thin-section and negative-staining electron microscopy confirms the absence of sarcoplasmic reticulum and mitochondrial contamination. 5/zm in diameter (Fig. 2). Relatively few contaminating transverse tubules are observed. Transverse tubules are observed in other fractions (especially in band 3 of the sucrose gradient, see Fig. IA), as indicated by characteristic morphology of the isolated transverse tubules and the higher content of nitrendipine receptor sites than for the sarcolemmal vesicle fraction (unpublished observations).

Conventional cuvettes and spectrofluorometers should not be used for the following reasons: (1) The maximum amount of Ca 2÷ released from a skinned skeletal muscle fiber with a diameter of 100/zm is of the order of I0 pmol/mm of fiber length. 1/zM increase in Ca 2÷ concentration even if a 10-mm-long skinned fiber is used. On the other hand, in a capillary with an internal diameter of 400/zm, several tens of a micromolar change in Ca :+ concentration can be produced. (2) Solution changes cannot be carried out rapidly using a conventional cuvette and large quantities of solutions containing fura-2 (which is rather expensive) are discarded.

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