Bublitz M. (Ed.) P-Type ATPases. Methods and Protocols

Humana Press, 2016 — 544p. — (Methods in Molecular Biology 1377) — ISBN: 978-1-4939-3179-8 (eBook), 978-1-4939-3178-1 (Hardcover).This volume provides a broad collection of protocols for many of the common experimental techniques used for the characterization of P-type ATPases. P-Type ATPases: Methods and Protocols provides comprehensive practical instructions for all researchers in the P-type ATPase field, from the protein biochemist to the mouse geneticist, covering the identification and classification, isolation, purification, in vitro characterization, knock-out studies, as well as crystallization and structural analysis. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, P-Type ATPases: Methods and Protocols aims to ensure successful results in the further study of this vital field of research.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 P_IB-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