Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.

Specialized trafficking systems in eukaryotic cells serve a critical role in partitioning intracellular proteins between the nucleus and cytoplasm. Cytoplasmic proteins (including chromatin remodeling enzymes and transcription factors) must gain access to the nucleus to exert their functions to prop...

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Autores principales:	Ki-Eun Park, H Dorota Inerowicz, Xin Wang, Yanfang Li, Stephanie Koser, Ryan A Cabot
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/028bd94dbec0485dba2c282f73d893e1
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spelling	oai:doaj.org-article:028bd94dbec0485dba2c282f73d893e12021-11-18T07:15:35ZIdentification of karyopherin α1 and α7 interacting proteins in porcine tissue.1932-620310.1371/journal.pone.0038990https://doaj.org/article/028bd94dbec0485dba2c282f73d893e12012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22720010/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Specialized trafficking systems in eukaryotic cells serve a critical role in partitioning intracellular proteins between the nucleus and cytoplasm. Cytoplasmic proteins (including chromatin remodeling enzymes and transcription factors) must gain access to the nucleus to exert their functions to properly program fundamental cellular events ranging from cell cycle progression to gene transcription. Knowing that nuclear import mediated by members of the karyopherin α family of transport receptors plays a critical role in regulating development and differentiation, we wanted to determine the identity of proteins that are trafficked by this karyopherin α pathway. To this end, we performed a GST pull-down assay using porcine orthologs of karyopherin α1 (KPNA1) and karyopherin α7 (KPNA7) and prey protein derived from porcine fibroblast cells and used a liquid chromatography and tandem mass spectrometry (LC-MS/MS) approach to determine the identity of KPNA1 and KPNA7 interacting proteins. Our screen revealed that the proteins that interact with KPNA1 and KPNA7 are generally nuclear proteins that possess nuclear localization signals. We further validated two candidate proteins from this screen and showed that they are able to be imported into the nucleus in vivo and also interact with members of the karyopherin α family of proteins in vitro. Our results also reveal the utility of using a GST pull-down approach coupled with LC-MS/MS to screen for protein interaction partners in a non-traditional model system.Ki-Eun ParkH Dorota InerowiczXin WangYanfang LiStephanie KoserRyan A CabotPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 6, p e38990 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ki-Eun Park H Dorota Inerowicz Xin Wang Yanfang Li Stephanie Koser Ryan A Cabot Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.
description	Specialized trafficking systems in eukaryotic cells serve a critical role in partitioning intracellular proteins between the nucleus and cytoplasm. Cytoplasmic proteins (including chromatin remodeling enzymes and transcription factors) must gain access to the nucleus to exert their functions to properly program fundamental cellular events ranging from cell cycle progression to gene transcription. Knowing that nuclear import mediated by members of the karyopherin α family of transport receptors plays a critical role in regulating development and differentiation, we wanted to determine the identity of proteins that are trafficked by this karyopherin α pathway. To this end, we performed a GST pull-down assay using porcine orthologs of karyopherin α1 (KPNA1) and karyopherin α7 (KPNA7) and prey protein derived from porcine fibroblast cells and used a liquid chromatography and tandem mass spectrometry (LC-MS/MS) approach to determine the identity of KPNA1 and KPNA7 interacting proteins. Our screen revealed that the proteins that interact with KPNA1 and KPNA7 are generally nuclear proteins that possess nuclear localization signals. We further validated two candidate proteins from this screen and showed that they are able to be imported into the nucleus in vivo and also interact with members of the karyopherin α family of proteins in vitro. Our results also reveal the utility of using a GST pull-down approach coupled with LC-MS/MS to screen for protein interaction partners in a non-traditional model system.
format	article
author	Ki-Eun Park H Dorota Inerowicz Xin Wang Yanfang Li Stephanie Koser Ryan A Cabot
author_facet	Ki-Eun Park H Dorota Inerowicz Xin Wang Yanfang Li Stephanie Koser Ryan A Cabot
author_sort	Ki-Eun Park
title	Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.
title_short	Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.
title_full	Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.
title_fullStr	Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.
title_full_unstemmed	Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.
title_sort	identification of karyopherin α1 and α7 interacting proteins in porcine tissue.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/028bd94dbec0485dba2c282f73d893e1
work_keys_str_mv	AT kieunpark identificationofkaryopherina1anda7interactingproteinsinporcinetissue AT hdorotainerowicz identificationofkaryopherina1anda7interactingproteinsinporcinetissue AT xinwang identificationofkaryopherina1anda7interactingproteinsinporcinetissue AT yanfangli identificationofkaryopherina1anda7interactingproteinsinporcinetissue AT stephaniekoser identificationofkaryopherina1anda7interactingproteinsinporcinetissue AT ryanacabot identificationofkaryopherina1anda7interactingproteinsinporcinetissue
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Identification of karyopherin α1 and α7 interacting proteins in porcine tissue.

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