Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4

Abstract Our recent studies of peptidylarginine deiminase 4 (PAD4) demonstrate that its non-catalytic Ca2+-binding sites play a crucial role in the assembly of the correct geometry of the enzyme. Here, we examined the folding mechanism of PAD4 and the role of Ca2+ ions in the folding pathway. Multip...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Yi-Liang Liu, Chien-Yun Lee, Yu-Ni Huang, Hui-Yi Chen, Guang-Yaw Liu, Hui-Chih Hung
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9aceff9d3ef9404389090cd44055a0a0
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:9aceff9d3ef9404389090cd44055a0a0
record_format	dspace
spelling	oai:doaj.org-article:9aceff9d3ef9404389090cd44055a0a02021-12-02T16:07:48ZProbing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 410.1038/s41598-017-02677-12045-2322https://doaj.org/article/9aceff9d3ef9404389090cd44055a0a02017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02677-1https://doaj.org/toc/2045-2322Abstract Our recent studies of peptidylarginine deiminase 4 (PAD4) demonstrate that its non-catalytic Ca2+-binding sites play a crucial role in the assembly of the correct geometry of the enzyme. Here, we examined the folding mechanism of PAD4 and the role of Ca2+ ions in the folding pathway. Multiple mutations were introduced into the calcium-binding sites, and these mutants were termed the Ca1_site, Ca2_site, Ca3_site, Ca4_site and Ca5_site mutants. Our data indicate that during the unfolding process, the PAD4 dimer first dissociates into monomers, and the monomers then undergo a three-state denaturation process via an intermediate state formation. In addition, Ca2+ ions assist in stabilizing the folding intermediate, particularly through binding to the Ca3_site and Ca4_site to ensure the correct and active conformation of PAD4. The binding of calcium ions to the Ca1_site and Ca2_site is directly involved in the catalytic action of the enzyme. Finally, this study proposes a model for the folding of PAD4. The nascent polypeptide chains of PAD4 are first folded into monomeric intermediate states, then continue to fold into monomers, and ultimately assemble into a functional and dimeric PAD4 enzyme, and cellular Ca2+ ions may be the critical factor governing the interchange.Yi-Liang LiuChien-Yun LeeYu-Ni HuangHui-Yi ChenGuang-Yaw LiuHui-Chih HungNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-14 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yi-Liang Liu Chien-Yun Lee Yu-Ni Huang Hui-Yi Chen Guang-Yaw Liu Hui-Chih Hung Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4
description	Abstract Our recent studies of peptidylarginine deiminase 4 (PAD4) demonstrate that its non-catalytic Ca2+-binding sites play a crucial role in the assembly of the correct geometry of the enzyme. Here, we examined the folding mechanism of PAD4 and the role of Ca2+ ions in the folding pathway. Multiple mutations were introduced into the calcium-binding sites, and these mutants were termed the Ca1_site, Ca2_site, Ca3_site, Ca4_site and Ca5_site mutants. Our data indicate that during the unfolding process, the PAD4 dimer first dissociates into monomers, and the monomers then undergo a three-state denaturation process via an intermediate state formation. In addition, Ca2+ ions assist in stabilizing the folding intermediate, particularly through binding to the Ca3_site and Ca4_site to ensure the correct and active conformation of PAD4. The binding of calcium ions to the Ca1_site and Ca2_site is directly involved in the catalytic action of the enzyme. Finally, this study proposes a model for the folding of PAD4. The nascent polypeptide chains of PAD4 are first folded into monomeric intermediate states, then continue to fold into monomers, and ultimately assemble into a functional and dimeric PAD4 enzyme, and cellular Ca2+ ions may be the critical factor governing the interchange.
format	article
author	Yi-Liang Liu Chien-Yun Lee Yu-Ni Huang Hui-Yi Chen Guang-Yaw Liu Hui-Chih Hung
author_facet	Yi-Liang Liu Chien-Yun Lee Yu-Ni Huang Hui-Yi Chen Guang-Yaw Liu Hui-Chih Hung
author_sort	Yi-Liang Liu
title	Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4
title_short	Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4
title_full	Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4
title_fullStr	Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4
title_full_unstemmed	Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4
title_sort	probing the roles of calcium-binding sites during the folding of human peptidylarginine deiminase 4
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/9aceff9d3ef9404389090cd44055a0a0
work_keys_str_mv	AT yiliangliu probingtherolesofcalciumbindingsitesduringthefoldingofhumanpeptidylargininedeiminase4 AT chienyunlee probingtherolesofcalciumbindingsitesduringthefoldingofhumanpeptidylargininedeiminase4 AT yunihuang probingtherolesofcalciumbindingsitesduringthefoldingofhumanpeptidylargininedeiminase4 AT huiyichen probingtherolesofcalciumbindingsitesduringthefoldingofhumanpeptidylargininedeiminase4 AT guangyawliu probingtherolesofcalciumbindingsitesduringthefoldingofhumanpeptidylargininedeiminase4 AT huichihhung probingtherolesofcalciumbindingsitesduringthefoldingofhumanpeptidylargininedeiminase4
_version_	1718384733419732992

Probing the Roles of Calcium-Binding Sites during the Folding of Human Peptidylarginine Deiminase 4

Ejemplares similares