Investigating the oxidative refolding mechanism of Cripto-1 CFC domain(44 visite) Iaccarino E, Sandomenico A, Corvino G, Foca G, Severino V, Russo R, Caporale A, Raimondo D, D'Abramo M, Alba J, Chambery A, Ruvo M
Impact factor: 4.784, Impact factor a 5 anni: 3.227
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Parole chiave: Cfc Domain, Cripto, Disulfide Bridges, Oxidative Folding,
*** IBB - CNR *** Dipartimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche (DISTABIF), via Vivaldi, 43, 80100 Caserta, Italy Istituto di Biostrutture e Bioimmagini, CNR and Centro Interuniversitario di Ricerca sui Peptidi Bioattivi (CIRPeB), via Mezzocannone 16, 80134 Napoli, Italy. Dipartimento di Medicina Molecolare, Sapienza Universita di Roma, 00161, Italy. Dipartimento di Chimica, Sapienza Universita di Roma, 00161, Italy. Anbition srl, via A. Manzoni, 1, 80123 Napoli, Italy. Electronic address: email@example.com.
Using a combined approach based on MS, enzyme digestion and advanced MD studies we have determined the sequential order of formation of the three disulfide bridges of the Cripto-1 CFC domain. The domain has a rare pattern of bridges and is involved in the recognition of several receptors. The bridge formation order is C1-C4, C3-C5, C2-C6, however formation of C1-C4 plays no roles for the formation of the others. Folding is driven by formation of the C3-C5 bridge and is supported by residues lying within the segment delimited by these cysteines. We indeed observe that variants CFC-W123A and CFC-DeltaC1/C4, where C1 and C4 are replaced by serines, are able to refold in the same time window as the wild type, while CFC-K132A and CFC-W134A are not. A variant where cysteines of the second and third bridge are mutated to serine, convert slowly to the monocyclic molecule. Data altogether support a mechanism whereby the Cripto-1 CFC domain refolds by virtue of long-range intramolecular interactions that involve residues close to cysteines of the second and third bridge. These findings are supported by the in silico study that shows how distant parts of the molecules come into contact on a long time scale.<br>
10 Records (10 escludendo Abstract e Conferenze). Impact factor totale: 29.734 (29.734 escludendo Abstract e Conferenze). Impact factor a 5 anni totale: 30.615 (30.615 escludendo Abstract e Conferenze).