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Promotion of PDGF-induced endothelial cell migration by phosphorylated VASP depends on PKA anchoring via AKAP

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Abstract

Vasodilator-stimulated phosphoprotein (VASP), an important substrate of PKA, plays a critical role in remodeling of actin cytoskeleton and actin-based cell motility. However, how PKA accurately transfers extracellular signals to VASP and then how phosphorylation of VASP regulates endothelial cell migration have not been clearly defined. Protein kinase A anchoring proteins (AKAPs) are considered to regulate intracellular-specific signal targeting of PKA via AKAP-mediated PKA anchoring. Thus, our study investigated the relationship among AKAP anchoring of PKA, PKA activity, and VASP phosphorylation, which is to clarify the exact role of VASP and its upstream regulatory mechanism in PKA-dependent migration. Our results show that chemotactic factor PDGF activated PKA, increased phosphorylation of VASP at Ser157, and enhanced ECV304 endothelial cell migration. However, phosphorylation site-directed mutation of VASP at Ser157 attenuated the chemotactic effect of PDGF on endothelial cells, suggesting phosphorylation of VASP at Ser157 promotes PKA-mediated endothelial cell migration. Furthermore, disrupting PKA anchoring to AKAP or PKA activity significantly attenuated the PKA activity, VASP phosphorylation, and subsequent cell migration. Meanwhile, disrupting PKA anchoring to AKAP abolished PDGF-induced lamellipodia formation and special VASP accumulation at leading edge of lamellipodia. These results indicate that PKA activation and PKA-mediated substrate responses in VASP phosphorylation and localization depend on PKA anchoring via AKAP in PDGF-induced endothelial cell migration. In conclusion, AKAP anchoring of PKA is an essential upstream event in regulation of PKA-mediated VASP phosphorylation and subsequent endothelial cell migration, which contributes to explore new methods for controlling endothelial cell migration related diseases and angiogenesis.

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Acknowledgments

This study was supported by grants from National Natural Science Foundation of China (Grant No. 30470680). Recombinant plasmids (VASP-WT and Ser157 mutant VASP-S157A) are kindly gifts from the Institute of Clinical Biochemistry and Pathobiochemistry, Wurzburg University, Germany.

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  1. Department of Pathophysiology, Medical College of Wuhan University, DongHu Road 115, Wuhan, Hubei Province, China

    Deling Zhang, Jingping Ouyang, Nian Wang, Yahui Zhang, Jinghua Bie & Yemin Zhang

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  1. Deling Zhang
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  2. Jingping Ouyang
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Correspondence to Jingping Ouyang.

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Zhang, D., Ouyang, J., Wang, N. et al. Promotion of PDGF-induced endothelial cell migration by phosphorylated VASP depends on PKA anchoring via AKAP. Mol Cell Biochem 335, 1–11 (2010). https://doi.org/10.1007/s11010-009-0234-y

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  • DOI: https://doi.org/10.1007/s11010-009-0234-y

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