The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85

SH2 domains are protein domains that modulate protein−protein interactions through a specific interaction with sequences containing phosphorylated tyrosines. In this work, we analyze the folding pathway of the C-terminal SH2 domain of the p85 regulatory subunit of the protein PI3K, which p...

Full description

Bibliographic Details
Main Authors:	Francesca Troilo, Francesca Malagrinò, Lorenzo Visconti, Angelo Toto, Stefano Gianni
Format:	Article
Language:	English
Published:	MDPI AG 2019-12-01
Series:	International Journal of Molecular Sciences
Subjects:	protein folding misfolding kinetics mutagenesis
Online Access:	https://www.mdpi.com/1422-0067/21/1/125

id	doaj-a47e24c6ecf842729ea60f384a03c2ac
record_format	Article
spelling	doaj-a47e24c6ecf842729ea60f384a03c2ac2020-11-25T01:49:49ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-12-0121112510.3390/ijms21010125ijms21010125The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85Francesca Troilo0Francesca Malagrinò1Lorenzo Visconti2Angelo Toto3Stefano Gianni4Istituto Pasteur—Fondazione Cenci Bolognetti, Dipartimento di Scienze Biochimiche “A. Rossi Fanelli”, 00161 Roma, ItalyIstituto Pasteur—Fondazione Cenci Bolognetti, Dipartimento di Scienze Biochimiche “A. Rossi Fanelli”, 00161 Roma, ItalyIstituto Pasteur—Fondazione Cenci Bolognetti, Dipartimento di Scienze Biochimiche “A. Rossi Fanelli”, 00161 Roma, ItalyIstituto Pasteur—Fondazione Cenci Bolognetti, Dipartimento di Scienze Biochimiche “A. Rossi Fanelli”, 00161 Roma, ItalyIstituto Pasteur—Fondazione Cenci Bolognetti, Dipartimento di Scienze Biochimiche “A. Rossi Fanelli”, 00161 Roma, ItalySH2 domains are protein domains that modulate protein−protein interactions through a specific interaction with sequences containing phosphorylated tyrosines. In this work, we analyze the folding pathway of the C-terminal SH2 domain of the p85 regulatory subunit of the protein PI3K, which presents a proline residue in a cis configuration in the loop between the βE and βF strands. By employing single and double jump folding and unfolding experiments, we demonstrate the presence of an on-pathway intermediate that transiently accumulates during (un)folding. By comparing the kinetics of folding of the wild-type protein to that of a site-directed variant of C-SH2 in which the proline was replaced with an alanine, we demonstrate that this intermediate is dictated by the peptidyl prolyl cis-trans isomerization. The results are discussed in the light of previous work on the effect of peptidyl prolyl cis-trans isomerization on folding events.https://www.mdpi.com/1422-0067/21/1/125protein foldingmisfoldingkineticsmutagenesis
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Francesca Troilo Francesca Malagrinò Lorenzo Visconti Angelo Toto Stefano Gianni
spellingShingle	Francesca Troilo Francesca Malagrinò Lorenzo Visconti Angelo Toto Stefano Gianni The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85 International Journal of Molecular Sciences protein folding misfolding kinetics mutagenesis
author_facet	Francesca Troilo Francesca Malagrinò Lorenzo Visconti Angelo Toto Stefano Gianni
author_sort	Francesca Troilo
title	The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85
title_short	The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85
title_full	The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85
title_fullStr	The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85
title_full_unstemmed	The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85
title_sort	effect of proline <i>cis</i>-<i>trans</i> isomerization on the folding of the c-terminal sh2 domain from p85
publisher	MDPI AG
series	International Journal of Molecular Sciences
issn	1422-0067
publishDate	2019-12-01
description	SH2 domains are protein domains that modulate protein−protein interactions through a specific interaction with sequences containing phosphorylated tyrosines. In this work, we analyze the folding pathway of the C-terminal SH2 domain of the p85 regulatory subunit of the protein PI3K, which presents a proline residue in a cis configuration in the loop between the βE and βF strands. By employing single and double jump folding and unfolding experiments, we demonstrate the presence of an on-pathway intermediate that transiently accumulates during (un)folding. By comparing the kinetics of folding of the wild-type protein to that of a site-directed variant of C-SH2 in which the proline was replaced with an alanine, we demonstrate that this intermediate is dictated by the peptidyl prolyl cis-trans isomerization. The results are discussed in the light of previous work on the effect of peptidyl prolyl cis-trans isomerization on folding events.
topic	protein folding misfolding kinetics mutagenesis
url	https://www.mdpi.com/1422-0067/21/1/125
work_keys_str_mv	AT francescatroilo theeffectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT francescamalagrino theeffectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT lorenzovisconti theeffectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT angelototo theeffectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT stefanogianni theeffectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT francescatroilo effectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT francescamalagrino effectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT lorenzovisconti effectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT angelototo effectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85 AT stefanogianni effectofprolineicisiitransiisomerizationonthefoldingofthecterminalsh2domainfromp85
_version_	1725004756647149568

The Effect of Proline <i>cis</i>-<i>trans</i> Isomerization on the Folding of the C-Terminal SH2 Domain from p85

Similar Items