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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| Format: | Article |
| Language: | English |
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Dove Medical Press
2008-08-01
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| Series: | Biologics: Targets & Therapy |
| Online Access: | http://www.dovepress.com/use-of-a-decoy-peptide-to-purify-p21-activated-kinase-1-in-cardiac-mus-a2006 |
| _version_ | 1819292606241177600 |
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| author | Yunbo Ke R John Solaro |
| author_facet | Yunbo Ke R John Solaro |
| author_sort | Yunbo Ke |
| collection | DOAJ |
| 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 |
| first_indexed | 2024-12-24T03:57:12Z |
| format | Article |
| id | doaj.art-c762d725c9d7433fac1d91caf4fce75a |
| institution | Directory Open Access Journal |
| issn | 1177-5475 1177-5491 |
| language | English |
| last_indexed | 2024-12-24T03:57:12Z |
| publishDate | 2008-08-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | Biologics: Targets & Therapy |
| spelling | doaj.art-c762d725c9d7433fac1d91caf4fce75a2022-12-21T17:16:25ZengDove Medical PressBiologics: Targets & Therapy1177-54751177-54912008-08-012008Issue 4903909Use of a decoy peptide to purify p21 activated kinase-1 in cardiac muscle and identification of ceramide-related activationYunbo KeR John SolaroYunbo 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 ceramidehttp://www.dovepress.com/use-of-a-decoy-peptide-to-purify-p21-activated-kinase-1-in-cardiac-mus-a2006 |
| spellingShingle | 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 Biologics: Targets & Therapy |
| title | 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_short | 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 |
| url | http://www.dovepress.com/use-of-a-decoy-peptide-to-purify-p21-activated-kinase-1-in-cardiac-mus-a2006 |
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