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Membrane Protein Structure Determination: Methods and by Jean-Jacques Lacapere

By Jean-Jacques Lacapere

Membrane proteins, representing approximately forty% of all proteins, are key parts of cells eager about many mobile strategies, but just a small variety of their constructions were decided. Membrane Protein constitution choice: equipment and Protocols provides many distinct innovations for membrane protein constitution selection used this present day via bringing jointly contributions from most sensible specialists within the box. Divided into 5 handy sections, the e-book covers a number of options to purify membrane proteins, ways to get 3 dimensional crystals and clear up the constitution by way of x-ray diffraction, probabilities to achieve structural details for a membrane protein utilizing electron microscopy observations, contemporary advances in nuclear magnetic resonance (NMR), and molecular modelling thoughts that may be used both to get membrane protein buildings or to maneuver from atomic constitution to a dynamic figuring out of a molecular functioning mechanism. Written within the hugely profitable tools in Molecular Biology™ sequence layout, chapters contain introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, effortlessly reproducible laboratory protocols, and tips about troubleshooting and warding off recognized pitfalls. complete and straightforward to exploit, Membrane Protein constitution choice: equipment and Protocols serves as an incredible reference for scientists trying to extra our wisdom of those very important and flexible proteins in addition to our total knowing of the complex global of mobilephone biology.

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Additional info for Membrane Protein Structure Determination: Methods and Protocols (Methods in Molecular Biology, Vol. 654)

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Protein Expr Purif 19:57–65 5. Brandolin G, Doussiere J, Gulik A, GulikKrywicki T, Lauquin GJM, Vignais PV (1980) Kinetic, binding and ultrastructural properties of the beef heart adenine nucleotide carrier protein after incorporation into phospholipid vesicles. Biochim Biophys Acta 592:592–614 6. De Marcos Lousa C, Trézéguet V, Dianoux A-C, Brandolin G, Lauquin GJ-M (2002) 7. 8. 9. 10. 11. 12. The human mitochondrial ADP/ATP carriers: kinetic properties and biogenesis of wild type and mutant proteins in the yeast S.

Anal Biochem 267:46–56 Mokus M, Kragh-Hansen U, Letellier P, le Maire M, Møller JV (1998) Construction and use of a detergent-sensitive electrode to measure dodecyl sulfate activity and binding. Anal Biochem 264:34–40 Eriks LR, Mayor JA, Kaplan RS (2003) A strategy for identification and quantification of detergents frequently used in the purification of membrane proteins. Anal Biochem 323:234–241 Papadopoulos V, Baraldi M, Guilarte TR, Knudsen TB, Lacapere J-J, Lindemann P, Noremberg MD, Nutt D, Weizman A, Zhang M-R, Gavish M (2006) Translocator protein (18€kDa): new nomenclature for the peripheral-type benzodiazepine receptor based on 18 Ostuni et al.

MRecTSPO eluted in DPC (round black symbols). Resting mRecTSPO stacked in the column is eluted with buffer A supplemented with 50€mg/mL SDS (square white symbols). Total amount of mRecTSPO eluted in both DPC and SDS (triangular white symbols) is almost constant and close to 9€mg. We suggested that a minimal concentration of 5€ mg/mL DPC was necessary to elute a stable mRecTSPO detergent complex, which did not precipitate. It suggests that when DPC/mRecTSPO ratio was too low, protein tends to precipitate.

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