Chapter title |
Implementation of the NMR CHEmical Shift Covariance Analysis (CHESCA): A Chemical Biologist’s Approach to Allostery
|
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Chapter number | 18 |
Book title |
Protein NMR
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Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7386-6_18 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7385-9, 978-1-4939-7386-6
|
Authors |
Stephen Boulton, Rajeevan Selvaratnam, Rashik Ahmed, Giuseppe Melacini, Boulton, Stephen, Selvaratnam, Rajeevan, Ahmed, Rashik, Melacini, Giuseppe |
Abstract |
Mapping allosteric sites is emerging as one of the central challenges in physiology, pathology, and pharmacology. Nuclear Magnetic Resonance (NMR) spectroscopy is ideally suited to map allosteric sites, given its ability to sense at atomic resolution the dynamics underlying allostery. Here, we focus specifically on the NMR CHEmical Shift Covariance Analysis (CHESCA), in which allosteric systems are interrogated through a targeted library of perturbations (e.g., mutations and/or analogs of the allosteric effector ligand). The atomic resolution readout for the response to such perturbation library is provided by NMR chemical shifts. These are then subject to statistical correlation and covariance analyses resulting in clusters of allosterically coupled residues that exhibit concerted responses to the common set of perturbations. This chapter provides a description of how each step in the CHESCA is implemented, starting from the selection of the perturbation library and ending with an overview of different clustering options. |
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