Preparation of Caveolin-1 for NMR Spectroscopy Experiments.
Related Articles Preparation of Caveolin-1 for NMR Spectroscopy Experiments.
Methods Mol Biol. 2020;2169:137-147
Authors: Plucinsky SM, Julien JA, Glover KJ
Abstract
Caveolin-1 is a 20.5*kDa integral membrane protein that is involved in a myriad of cellular processes including signal transduction, relieving mechano-stresses on the cell, endocytosis, and most importantly caveolae formation. As a consequence, there is intense interest in characterizing caveolin-1 structurally. Out of the many available structural techniques, nuclear magnetic resonance (NMR) spectroscopy is particularly well suited to investigations on integral membrane proteins like caveolin-1 that have significant unstructured regions and unusual topologies. However, the technique requires relatively large amounts of protein (i.e. concentrations in the 0.5-5*mM range), and obtaining these amounts can be difficult especially for highly hydrophobic membrane proteins such as caveolin-1. Herein, we describe a robust protocol for the preparation of caveolin-1 for structural studies using NMR.
PMID: 32548826 [PubMed - in process]
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