Titlebook: BCL-2 Family Proteins; Methods and Protocol Evripidis Gavathiotis Book 2019 Springer Science+Business Media, LLC, part of Springer Nature 2

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BCL-2 Protein Family Interaction Analysis by Nuclear Magnetic Resonance Spectroscopy,ll molecule binding. NMR has been used extensively in the investigation of BCL-2 family proteins revealing the structure of key family members, identifying binding partners and interaction sites, and screening small molecule modulators. In this chapter we discuss the application of NMR to identify i

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Reconstitution and Characterization of BCL-2 Family Proteins in Lipid Bilayer Nanodiscs,drugs. Because their functions are intimately connected with intracellular membranes, it is important to perform structural and activity studies in precisely characterized samples that include phospholipids and capture the features of the native physiological environment as closely as possible. NMR

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Rapid Imaging of BCL-2 Family Interactions in Live Cells Using FLIM-FRET,to detect Förster resonance energy transfer (FRET) between two fluorescent-fusion proteins in live cells. FLIM-FRET has been used to detect specific interactions and their disruption, for Bcl-2 family proteins. To date, this has been possible only in low throughput, due to the time required for seri

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Mitochondrial Isolation and Real-Time Monitoring of MOMP,subset of MOMP regulators. Here we describe the method to isolate, JC-1-label, and purify mouse liver mitochondria and subsequently describe how to utilize the JC-1-labeled mitochondria for real-time MOMP measurements.

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Methods to Probe Calcium Regulation by BCL-2 Family Members,ortant function is the release of calcium from the endoplasmic reticulum (ER), which critically contributes to the process of apoptosis. Here, we describe a protocol to measure calcium levels in the ER, mitochondria, and cytosol, with specific consideration of BCL-2 family biology.

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