P-Type ATPases : Methods and Protocols - Bublitz, Maike; SpringerLink (Online service) (New York, NY : Springer New York : Imprint: Humana, 2016)
An Introduction to P-type ATPase research -- Purification of Na,K-ATPase from Pig Kidney -- Preparation of Ca2+-ATPase1a Enzyme from Rabbit Sarcoplasmic Reticulum -- Isolation of H+/K+-ATPase-enriched Membrane Fraction from Pig Stomachs -- Overproduction of PiB-type ATPases -- Coordinated Overexpression in Yeast of a P4-ATPase and its Associated Cdc50 Subunit: The Case of the Drs2p/Cdc50p Lipid Flippase Complex -- The Plasma Membrane Ca2+-ATPase: Purification by Calmodulin Affinity Chromatography, and Reconstitution of the Purified Protein -- Expression of Na,K-ATPase and H,K-ATPase Isoforms With The Baculovirus Expression System -- Time-dependent Protein Thermostability Assay -- Colorimetric Assays of Na,K-ATPase -- ATPase Activity Measurements by an Enzyme-Coupled Spectrophotometric Assay -- Antimony-Phosphomolybdate ATPase Assay -- ATPase Activity Measurements using radiolabeled ATP -- Assaying P-type ATPases Reconstituted in Liposomes -- Coupling ratio for Ca2+ Transport by Calcium Oxalate Precipitation -- Calcium Uptake in Crude Tissue Preparation -- Measuring H+ Pumping and Membrane Potential Formation in Sealed Membrane Vesicle Systems -- Assay of Flippase Activity in Proteoliposomes using Fluorescent Lipid Derivatives -- The use of Metal Fluoride Compounds as Phosphate Analogs for Understanding the Structural Mechanism in P-type ATPases -- Phosphorylation/Dephosphorylation Assays -- Tryptophan Fluorescence Changes Related to Ca2+-ATPase Function -- Determination of the ATP Affinity of the Sarcoplasmic Reticulum Ca2+-ATPase by Competitive Inhibition of [g-32P]TNP-8N3-ATP Photolabeling. 23. Ca2+ Binding and Transport Studied with Ca2+/EGTA Buffers and 45Ca2+ 24. Assay of Copper Transfer and Binding to P1B-ATPases 25. Voltage Clamp Fluorometry of P-Type ATPases 26. Electrophysiological Measurements on Solid Supported Membranes 27. Electrophysiological Characterization of Na,K-ATPases Expressed in Xenopus laevis Oocytes Using Two-Electrode Voltage Clamping 28. Functional Studies of Na+,K+-ATPase using Transfected Cell Cultures 29. HPLC Neurotransmitter Analysis 30. Behavior Test Relevant to a2/a3Na+/K+-ATPase Gene Modified Mouse Models 31. Zebrafish Whole-mount in situ Hybridization Followed by Sectioning 32. Whole-mount Immunohistochemistry for Anti-F59 in Zebrafish Embryos (1-5 days Post Fertilization (dpf)) Canan Doganli 33. Cell-based Lipid Flippase Assay Employing Fluorescent Lipid Derivatives 34. Transient Expression of P-type ATPases in Tobacco Epidermal Cells 35. Lipid Exchange by Ultracentrifugation 36. Reconstitution of Na+, K+-ATPase in Nanodiscs 37. Crystallization of P-type ATPases by the High Lipid-detergent (HiLiDe) Method 38. Two-dimensional Crystallization of the Ca2+-ATPase for Electron Crystallography 39. Two-dimensional Crystallization of Gastric H+,K+-ATPase for Structural Analysis By Electron Crystallography 40. MD Simulations of P-type ATPases in a Lipid Bilayer System 41. Computational Classification of P-Type ATPases 42. Molecular Modeling of Fluorescent SERCA Biosensors 43. How to Compare, Analyze and Morph Between Crystal Structures of Different Conformations: The P-type ATPase Example Jesper L .
Keywords: Cell membranes .; Membrane Biology