Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation

Yunbo Ke, R John SolaroDepartment of Physiology and Biophysics, Center for Cardiovascular Research, University of Illinois at Chicago, Chicago, IL, USAAbstract: The p21 activated kinase-1 (Pak1) is a serine-threonine protein kinase directly activated by Cdc42 and Rac1. In cardiac myocytes, Pak1 acti...

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Autores principales:	Yunbo Ke, R John Solaro
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2008
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/c762d725c9d7433fac1d91caf4fce75a
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spelling	oai:doaj.org-article:c762d725c9d7433fac1d91caf4fce75a2021-12-02T06:53:30ZUse of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation1177-54751177-5491https://doaj.org/article/c762d725c9d7433fac1d91caf4fce75a2008-08-01T00:00:00Zhttp://www.dovepress.com/use-of-a-decoy-peptide-to-purify-p21-activated-kinase-1-in-cardiac-mus-a2006https://doaj.org/toc/1177-5475https://doaj.org/toc/1177-5491Yunbo Ke, R John SolaroDepartment of Physiology and Biophysics, Center for Cardiovascular Research, University of Illinois at Chicago, Chicago, IL, USAAbstract: The p21 activated kinase-1 (Pak1) is a serine-threonine protein kinase directly activated by Cdc42 and Rac1. In cardiac myocytes, Pak1 activation leads to dephosphorylation of cTnI and C-protein through upregulation of phosphatase-2A (PP2A). Pak1 activity is directly correlated with its autophosphorylation, which occurs upon binding to the small GTPases and to some small organic molecules as well. In this report, we describe a novel method for rapid purification of endogenous Pak1 from bovine ventricle muscle. The method is simple and easy to carry out. The purified Pak1 demonstrated autophosphorylation in vitro that was enhanced by D-erythro-sphingosine-1, N-acetyl-D-erythro-sphingosine (C2-ceramide), and N-hexanoyl-D-erythro-sphingosine (C6-ceramide). Dihydro-L-threo-sphingosine (saphingol) also had some effect on Pak1 autophosphorylation. The method we developed provides a useful tool to study Pak1 activity and regulation in the heart. Moreover, our results indicate a potential role of the sphingolipids as unique signaling molecules inducing a direct activation of Pak1 that may modulate different cardiac functions.Keywords: pak1, heart muscle, purification, C2 ceramide, C6 ceramide Yunbo KeR John SolaroDove Medical PressarticleMedicine (General)R5-920ENBiologics: Targets & Therapy, Vol 2008, Iss Issue 4, Pp 903-909 (2008)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Yunbo Ke R John Solaro Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation
description	Yunbo Ke, R John SolaroDepartment of Physiology and Biophysics, Center for Cardiovascular Research, University of Illinois at Chicago, Chicago, IL, USAAbstract: The p21 activated kinase-1 (Pak1) is a serine-threonine protein kinase directly activated by Cdc42 and Rac1. In cardiac myocytes, Pak1 activation leads to dephosphorylation of cTnI and C-protein through upregulation of phosphatase-2A (PP2A). Pak1 activity is directly correlated with its autophosphorylation, which occurs upon binding to the small GTPases and to some small organic molecules as well. In this report, we describe a novel method for rapid purification of endogenous Pak1 from bovine ventricle muscle. The method is simple and easy to carry out. The purified Pak1 demonstrated autophosphorylation in vitro that was enhanced by D-erythro-sphingosine-1, N-acetyl-D-erythro-sphingosine (C2-ceramide), and N-hexanoyl-D-erythro-sphingosine (C6-ceramide). Dihydro-L-threo-sphingosine (saphingol) also had some effect on Pak1 autophosphorylation. The method we developed provides a useful tool to study Pak1 activity and regulation in the heart. Moreover, our results indicate a potential role of the sphingolipids as unique signaling molecules inducing a direct activation of Pak1 that may modulate different cardiac functions.Keywords: pak1, heart muscle, purification, C2 ceramide, C6 ceramide
format	article
author	Yunbo Ke R John Solaro
author_facet	Yunbo Ke R John Solaro
author_sort	Yunbo Ke
title	Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation
title_short	Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation
title_full	Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation
title_fullStr	Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation
title_full_unstemmed	Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation
title_sort	use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation
publisher	Dove Medical Press
publishDate	2008
url	https://doaj.org/article/c762d725c9d7433fac1d91caf4fce75a
work_keys_str_mv	AT yunboke useofadecoypeptidetopurifyp21activatedkinase1incardiacmuscleandidentificationofceramiderelatedactivation AT rjohnsolaro useofadecoypeptidetopurifyp21activatedkinase1incardiacmuscleandidentificationofceramiderelatedactivation
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Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activation

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