Expression, activation, and subcellular localization of the Rap1 GTPase in cord blood-derived human megakaryocytes

Alessandra Balduini, Alessandro Pecci, Paolo Lova, Nicoletta Arezzi, Carmela Marseglia, Federica Bellora, Cesare Perotti, Cesare Balduini, Carlo L. Balduini, Mauro Torti

Research output: Contribution to journalArticle

Abstract

The expression of the small GTPase Rap1 in human megakaryocytes (MKs) differentiated from cord blood (CB)-derived progenitors was investigated. High levels of Rap1 were detected in the majority of mature megakaryocytes independently of days of culture, while a very low percentage of immature megakaryocytes was found to express a small amount of the protein. Rap1 was predominantly detected on internal α-granule but not on the plasma membrane. By contrast, CD41 was clearly present on the peripheral plasma membrane, although it also displayed an intracellular localization similar to that of Rap1. Upon thrombin stimulation, both Rap1 and CD41 translocated to the periphery of the cell. At the opposite, RhoA GTPase and glycoprotein Ibα were predominantly located at the plasma membrane and did not undergo relocation upon thrombin stimulation. Thrombin induced a dose- and time-dependent activation of Rap1 in mature megakaryocytes. By using a confocal microscopy approach with a specific probe, active Rap1 was detected exclusively at the peripheral plasma membrane. These results demonstrate that expression of Rap1 occurs during maturation rather than differentiation of megakaryocytes from cord blood progenitor cells. Moreover, we demonstrate that thrombin-activated Rap1 is exclusively localized at the peripheral plasma membrane.

Original languageEnglish
Pages (from-to)84-93
Number of pages10
JournalExperimental Cell Research
Volume300
Issue number1
DOIs
Publication statusPublished - Oct 15 2004

Keywords

  • Activation
  • Differentiation
  • Megakaryocytes
  • Rap1
  • Thrombin

ASJC Scopus subject areas

  • Cell Biology

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