EF-hand calcium sensors respond structurally to changes in intracellular Ca2+ concentration, triggering diverse cellular responses and resulting in broad interactomes. Despite impressive advances in decoding their structure-function relationships, the folding mechanism of neuronal calcium sensors is still elusive. We used single-molecule optical tweezers to study the folding mechanism of the human neuronal calcium sensor 1 (NCS1). Two intermediate structures induced by Ca2+ binding to the EF-hands were observed during refolding. The complete folding of the C domain is obligatory for the folding of the N domain, showing striking interdomain dependence. Molecular dynamics results reveal the atomistic details of the unfolding process and rationalize the different domain stabilities during mechanical unfolding. Through constant-force experiments and hidden Markov model analysis, the free energy landscape of the protein was reconstructed. Our results emphasize that NCS1 has evolved a remarkable complex interdomain cooperativity and a fundamentally different folding mechanism compared to structurally related proteins. © 2013 Elsevier Ltd.

Heidarsson P.O., Otazo M.R., Bellucci L., Mossa A., Imparato A., Paci E., et al. (2013). Single-molecule folding mechanism of an EF-hand neuronal calcium sensor. STRUCTURE, 21(10), 1812-1821 [10.1016/j.str.2013.07.022].

Single-molecule folding mechanism of an EF-hand neuronal calcium sensor

Paci E.;
2013

Abstract

EF-hand calcium sensors respond structurally to changes in intracellular Ca2+ concentration, triggering diverse cellular responses and resulting in broad interactomes. Despite impressive advances in decoding their structure-function relationships, the folding mechanism of neuronal calcium sensors is still elusive. We used single-molecule optical tweezers to study the folding mechanism of the human neuronal calcium sensor 1 (NCS1). Two intermediate structures induced by Ca2+ binding to the EF-hands were observed during refolding. The complete folding of the C domain is obligatory for the folding of the N domain, showing striking interdomain dependence. Molecular dynamics results reveal the atomistic details of the unfolding process and rationalize the different domain stabilities during mechanical unfolding. Through constant-force experiments and hidden Markov model analysis, the free energy landscape of the protein was reconstructed. Our results emphasize that NCS1 has evolved a remarkable complex interdomain cooperativity and a fundamentally different folding mechanism compared to structurally related proteins. © 2013 Elsevier Ltd.
2013
Heidarsson P.O., Otazo M.R., Bellucci L., Mossa A., Imparato A., Paci E., et al. (2013). Single-molecule folding mechanism of an EF-hand neuronal calcium sensor. STRUCTURE, 21(10), 1812-1821 [10.1016/j.str.2013.07.022].
Heidarsson P.O.; Otazo M.R.; Bellucci L.; Mossa A.; Imparato A.; Paci E.; Corni S.; Di Felice R.; Kragelund B.B.; Cecconi C.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/885098
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