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- Title
Resolution of Stepwise Cooperativities of Copper Binding by the Homotetrameric Copper-Sensitive Operon Repressor (CsoR): Impact on Structure and Stability.
- Authors
Jacobs, Alexander D.; Chang, Feng‐Ming James; Morrison, Lindsay; Dilger, Jonathan M.; Wysocki, Vicki H.; Clemmer, David E.; Giedroc, David P.
- Abstract
The cooperativity of ligand binding is central to biological regulation and new approaches are needed to quantify these allosteric relationships. Herein, we exploit a suite of mass spectrometry (MS) experiments to provide novel insights into homotropic Cu-binding cooperativity, gas-phase stabilities and conformational ensembles of the D2-symmetric, homotetrameric copper-sensitive operon repressor (CsoR) as a function of CuI ligation state. CuI binding is overall positively cooperative, but is characterized by distinct ligation state-specific cooperativities. Structural transitions occur upon binding the first and fourth CuI, with the latter occurring with significantly higher cooperativity than previous steps; this results in the formation of a holo-tetramer that is markedly more resistant than apo-, and partially ligated CsoR tetramers toward surface-induced dissociation (SID).
- Subjects
LIGAND binding (Biochemistry); PHYSIOLOGICAL control systems; STEPWISE reactions (Chemistry); MASS spectrometry; OPERONS
- Publication
Angewandte Chemie International Edition, 2015, Vol 54, Issue 43, p12795
- ISSN
1433-7851
- Publication type
Article
- DOI
10.1002/anie.201506349