Abstract
Pleiotrophin (PTN) stimulates endothelial cell migration through binding to receptor protein tyrosine phosphatase beta/zeta (RPTPβ/ζ) and ανβ3 integrin. Screening for proteins that interact with RPTPβ/ζ and potentially regulate PTN signaling, through mass spectrometry analysis, identified cyclin-dependent kinase 5 (CDK5) activator p35 among the proteins displaying high sequence coverage. Interaction of p35 with the serine/threonine kinase CDK5 leads to CDK5 activation, known to be implicated in cell migration. Protein immunoprecipitation and proximity ligation assays verified p35-RPTPβ/ζ interaction and revealed the molecular association of CDK5 and RPTPβ/ζ. In endothelial cells, PTN activates CDK5 in an RPTPβ/ζ- and phosphoinositide 3-kinase (PI3K)-dependent manner. On the other hand, c-Src, ανβ3 and ERK1/2 do not mediate the PTN-induced CDK5 activation. Pharmacological and genetic inhibition of CDK5 abolished PTN-induced endothelial cell migration, suggesting that CDK5 mediates PTN stimulatory effect. A new pyrrolo[2,3-α]carbazole derivative previously identified as a CDK1 inhibitor, was found to suppress CDK5 activity and eliminate PTN stimulatory effect on cell migration, warranting its further evaluation as a new CDK5 inhibitor. Collectively, our data reveal that CDK5 is activated by PTN, in an RPTPβ/ζ-dependent manner, regulates PTN-induced cell migration and is an attractive target for the inhibition of PTN pro-angiogenic properties.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism
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Amino Acid Sequence
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Animals
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Carbazoles / pharmacology
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Carrier Proteins / pharmacology*
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Cell Cycle Proteins / genetics
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Cell Cycle Proteins / metabolism
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Cell Line, Tumor
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Cell Movement / drug effects*
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Cyclin-Dependent Kinase 5 / antagonists & inhibitors
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Cyclin-Dependent Kinase 5 / genetics*
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Cyclin-Dependent Kinase 5 / metabolism
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Cytokines / genetics
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Cytokines / metabolism
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Cytokines / pharmacology*
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Gene Expression Regulation
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Guanine / analogs & derivatives
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Guanine / pharmacology
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Human Umbilical Vein Endothelial Cells / cytology
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Human Umbilical Vein Endothelial Cells / drug effects*
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Human Umbilical Vein Endothelial Cells / metabolism
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Humans
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Integrin alphaVbeta3 / genetics
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Integrin alphaVbeta3 / metabolism
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Isoenzymes / genetics
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Isoenzymes / metabolism
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Isoenzymes / pharmacology
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Neuroglia / drug effects
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Neuroglia / metabolism
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Neuroglia / pathology
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Phosphatidylinositol 3-Kinases / genetics
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Phosphatidylinositol 3-Kinases / metabolism
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Phosphoinositide-3 Kinase Inhibitors
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Protein Binding / drug effects
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Protein Kinase Inhibitors / pharmacology
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Rats
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Receptor-Like Protein Tyrosine Phosphatases, Class 5 / genetics*
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Receptor-Like Protein Tyrosine Phosphatases, Class 5 / metabolism
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Roscovitine / pharmacology
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Signal Transduction
Substances
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Adaptor Proteins, Signal Transducing
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CDCA5 protein, human
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Carbazoles
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Carrier Proteins
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Cell Cycle Proteins
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Cytokines
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Integrin alphaVbeta3
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Isoenzymes
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NU2058
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Phosphoinositide-3 Kinase Inhibitors
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Protein Kinase Inhibitors
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Roscovitine
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pleiotrophin
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Guanine
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Cyclin-Dependent Kinase 5
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CDK5 protein, human
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Receptor-Like Protein Tyrosine Phosphatases, Class 5