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Thromb Haemost
DOI: 10.1055/a-2689-7785
Cellular Haemostasis and Platelets

EFCAB13 is a Novel Age-Dependent Promotor of SOCE in Blood Platelets and Megakaryoblastic MEG-01 Cells

Authors

  • Pedro C. Redondo 

    1   Department of Physiology (FICELL), University of Extremadura, Cáceres, Spain

  • María P. Granados

    2   Pharmacy Unit, Extremadura County Health Service, Cáceres, Spain

  • Esperanza Fernández

    3   Blood Donation Center, Extremadura County Health Service, Mérida, Spain

  • Miguel-A Collado-Pérez

    1   Department of Physiology (FICELL), University of Extremadura, Cáceres, Spain

  • Raúl Teruel-Montoya

    4   Centro de Bioquímica y Genética Clínica. Hospital Clínico Universitario Virgen de la Arrixaca. Instituto Murciano de investigación Biosanitaria, El Palmar, Murcia, Spain

  • Francisca Ferrer-Marín

    5   Fundación Jimenez Díaz (IIS-FJD) CIBERER, IMID, UCAM, Murcia, Spain

  • Alejandro Berna-Erro 

    6   Department of Biochemistry and molecular Biology. Miguel Hernández University, Alicante, Spain

Funding The Spanish government (MICIN) supported this work through a Grant PID2022-136279-C21 funded by MCIN/AEI/10.13039/501100011033 and ERDF A way of making Europe, and by the Junta-Extremadura-FEDER research grants (IB18020 and GR21008). R.T.-M. and F.F.-M. were supported by Instituto de Salud Carlos III and FEDER (PI18/0316 and PI21/0347).
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Abstract

Background

Intracellular Ca2+ homeostasis is essential to regulate the molecular mechanisms underlying platelet physiology and aggregation. Store-operated Ca2+ entry (SOCE) is the main mechanism of extracellular Ca2+ influx in platelets and it has been involved in platelet aggregation.

Material and Methods

We reported the function of the uncharacterized protein EFCAB13 as a positive regulator of SOCE in megakaryoblastic cells and platelets from either mothers and their newborns. Alternatively, MEG-01 and HEK293 cells were used as sourrogated of platelets to perform changes in the expression of EFCAB13, which was ensured by immunoprecipitations and Western Blotting techniques. Once EFCAB13 was genetically modifies and to achive its role in Ca2+ homeostasis we used fluorescence microscope under single cells configuration. Finally, MEG-01 maturation was assay using flow-cytometry and specific fluorescent antibodies or cell cycle dies. Intracellular distribution and protein interaction was also evaluated by confocal microscopy.

Results

EFCAB13 is underexpressed in neonatal platelets as compared with adults, indicating an age-dependent expression. Silencing of EFCAB13 expression reduced the colocalization of STIM1 with Orai1 and, subsequently, impairs SOCE in megakaryoblastic MEG-01 cells stimulated with thapsigargin (TG). Coimmunoprecipitation experiments showed that EFCAB13 interacts with Orai1, and colocalization between both proteins increases during TG-evoked SOCE. Indeed, EFCAB13 localized nearby the ER during TG-evoked SOCE activation. Finally, PMA-induced MEG-01 maturation was altered in EFCAB13-silenced cells.

Conclusion

Altogether, we conclude that EFCAB13 binds to Orai1 to stabilize and regulate the STIM1/Orai1 complex formation, which is necessary to support SOCE in adults. However, its function is less important in neonatal platelets. This regulatory function may also be important for megakaryocytic physiology during maturation. Therefore, EFCAB13 could be an interesting target for specific treatment of maternal hemostatic disorders during pregnancy.

Keywords

EFCAB13 - STIM1 - Orai1 - SOCE - platelets

Data Availability Statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Please, contact pcr@unex.es or alejandrobe@unex.es for more details and information.


Ethical Approval Statement

This study followed the Helsinki's Declaration guidelines and the mandatory of the Ethical committees of the University of Extremadura and the Junta of Extremadura Health Service.


Authors' Contribution

P.C.R., and A.B.-E. contributed to conceptualization. E.F. and M.P.G. contributed to blood sample collection, and managing. P.C.R., A.B-E., R.T-M., and M.A.C-P. contributed to investigation. P.C.R. and A.B-E. contributed to validation. P.C.R. and A.B-E. contributed to supervision. A.B-E. contributed to writing the original manuscript. P.C.R., M.P.G., R.T-M., and F.F-M. contributed to manuscript editing. P.C.R. contributed to funding acquisition. P.C.R. contributed to project administration. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.


Supplementary Material



Publication History

Received: 07 April 2025

Accepted: 25 August 2025

Article published online:
19 September 2025

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