Download e-book for iPad: Biomembranes Part Q: ATP-Driven Pumps and Related Transport: by Sidney Fleischer, Becca Fleischer

By Sidney Fleischer, Becca Fleischer

ISBN-10: 0121820580

ISBN-13: 9780121820589

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

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

Sample text

24 C a 2+ FLUXES AND REGULATION [2] buffer in the assay solution. When this dye is excited by 340 nm ultraviolet light, the fluorescence intensity increases 3-fold upon binding of Ca 2+. For our experiments, we use a glass capillary as a cuvette and a fluorescence microscope equipped with a photomultiplier tube as a fluorometer. 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.

The same series of solution exchanges is made without any releasing solution for the controls. The result of experiments is presented as a value relative to that of the controls in Fig. 2B. The amount of Ca 2÷ in the control runs is taken as 100%. Figure 2B shows that the time courses are roughly exponential, and, therefore, the rate constant of the decline can be taken as the index of magnitude of Ca 2÷ release under these conditions. Essentially the same procedures can be followed for amphibian and mammalian (or avian) muscles.

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. This problem can be circumvented if a capillary cuvette is used. Using silk filaments, skinned fiber, 5 mm in length, is tied to a resincoated tungsten wire with a diameter of 100/xm. It is then inserted into and fixed in a glass capillary (internal diameter 400 ~m, length 32 mm) and mounted on the stage of a microscope. Both ends of the capillary are linked to silicone tubing.

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


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