By Reinhard Bolli, Clemens Broger, Angelo Azzi (auth.), Professor Dr. Angelo Azzi, Professor Dr. Lanfranco Masotti, Professor Dr. Arnaldo Vecli (eds.)
This quantity is the 3rd of a chain on Membrane Proteins and, just like the pre ceding manuals, is the results of a global complex path entitled Isolation and Characterization of Membrane Proteins: Biochemical and Bio actual features subsidized via the Federation of eu Biochemical So cieties (FEBS) and the Italian examine Council (CNR). The luck of the path and the continual improvement within the box of membrane biology has triggered me to submit additionally hence the protocols of the experiments that have been performed by means of the scholars. the scholars were capable not just to accomplish the experiments pub lished during this handbook with out aid from the teachers, but additionally to signify advancements, that have been included within the released model. Care has been taken in making the making plans and the execution of the ex periments so simple as attainable, via directory intimately the entire helpful items of kit, try tubes, pipettes, chemical substances, and so on. whilst the intro duction and the "philosophy" were constrained to the basic, as additionally the references, purely these having been indexed which could assist in a greater less than status of the rules and of the organic heritage of a given experi ment.
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Additional resources for Membrane Proteins: Isolation and Characterization
Sample text
00 During the experiment always keep the lipid samples under a nitrogen atmosphere and minimize exposure of the CHCl3 solutions oflipid to light. Open the vial of PS in CHCl3, keeping a nitrogen gas stream flowing over the vial. Transfer this solution to a 5-ml volumetric flask and make up to volume with CHCl3. If there is too much solvent, evaporate the excess using the nitrogen stream. Slowly bubble nitrogen gas through this stock solution for 10 min, keeping the volume constant by the addition of CHCl3.
A partly purified preparation of the enzyme (Rao and Keister 1978) has also been shown to act as a PPi-dependent electric generator (Konrashin et al. 1980), when incorporated into a planar phospholipid membrane. The PPase behaves like an integral membrane protein, spanning the entire chromatophore membrane. Earlier attempts to solubilize and purify this enzyme have only had very limited success (Rao and Keister 1978). Recently we have been able to obtain an apparently pure enzyme with reasonable stability in satisfactory yield (Nyren et a1.
The PPase behaves like an integral membrane protein, spanning the entire chromatophore membrane. Earlier attempts to solubilize and purify this enzyme have only had very limited success (Rao and Keister 1978). Recently we have been able to obtain an apparently pure enzyme with reasonable stability in satisfactory yield (Nyren et a1. 1984), and giving us the possibility to eventually perform a closer characterization of this protein, a characterization which, due to the analogy between the PPase and the ATPase and the apparent comparative simplicity of the former, may also provide closer insight into fundamental aspects of the functioning of coupling factor proteins in general.