SH2 domains are protein domains that mediate protein-protein interaction through the recognition and binding of specific sequences containing phosphorylated tyrosines. The p85 protein is the regulatory subunit of the heterodimeric enzyme PI3K, an important enzyme involved in several molecular pathways. In this work we characterize the folding kinetics of the NSH2 domain of p85. Our data clearly reveal peculiar folding kinetics, characterized by an apparent mismatch between the observed folding and unfolding kinetics. Taking advantage of double mixing stopped flow experiments and site directed mutagenesis we demonstrate that such behavior is due to the cis/trans isomerization of the peptide bond between D73 and P74, being in a cis conformation in the native protein. Our data are discussed in comparison with previous works on the folding of other SH2 domains.

The kinetics of folding of the NSH2 domain from p85 / Visconti, Lorenzo; Malagrinò, Francesca; Toto, Angelo; Gianni, Stefano. - In: SCIENTIFIC REPORTS. - ISSN 2045-2322. - 9:1(2019), p. 4058. [10.1038/s41598-019-40480-2]

The kinetics of folding of the NSH2 domain from p85

Visconti, Lorenzo;Malagrinò, Francesca;Toto, Angelo
;
Gianni, Stefano
2019

Abstract

SH2 domains are protein domains that mediate protein-protein interaction through the recognition and binding of specific sequences containing phosphorylated tyrosines. The p85 protein is the regulatory subunit of the heterodimeric enzyme PI3K, an important enzyme involved in several molecular pathways. In this work we characterize the folding kinetics of the NSH2 domain of p85. Our data clearly reveal peculiar folding kinetics, characterized by an apparent mismatch between the observed folding and unfolding kinetics. Taking advantage of double mixing stopped flow experiments and site directed mutagenesis we demonstrate that such behavior is due to the cis/trans isomerization of the peptide bond between D73 and P74, being in a cis conformation in the native protein. Our data are discussed in comparison with previous works on the folding of other SH2 domains.
2019
protein folding; stopped-flow; folding kinetics; proline isomerization
01 Pubblicazione su rivista::01a Articolo in rivista
The kinetics of folding of the NSH2 domain from p85 / Visconti, Lorenzo; Malagrinò, Francesca; Toto, Angelo; Gianni, Stefano. - In: SCIENTIFIC REPORTS. - ISSN 2045-2322. - 9:1(2019), p. 4058. [10.1038/s41598-019-40480-2]
File allegati a questo prodotto
File Dimensione Formato  
Visconti_The kinetics_2019.pdf

accesso aperto

Tipologia: Documento in Post-print (versione successiva alla peer review e accettata per la pubblicazione)
Licenza: Creative commons
Dimensione 1.06 MB
Formato Adobe PDF
1.06 MB Adobe PDF

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1253355
Citazioni
  • ???jsp.display-item.citation.pmc??? 4
  • Scopus 9
  • ???jsp.display-item.citation.isi??? 9
social impact