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Human umbilical vein endothelial cells secrete proteins via conventional and unconventional pathways, with 12.6% using unconventional cytoplasmic protein secretion (UCPS) for immune response and cell function.

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Area of Science:

  • Cell Biology
  • Immunology
  • Proteomics

Background:

  • Endothelial cells initiate innate immunity via pattern recognition receptors like Toll-like receptor 4 (TLR4).
  • Protein secretion occurs via classical ER-Golgi-dependent or unconventional ER-Golgi-independent pathways (UCPS).

Purpose of the Study:

  • To systematically characterize the human umbilical vein-derived endothelial cell (HUVEC) secretome.
  • To determine the contribution of UCPS to protein secretion in HUVECs, especially after TLR4 stimulation.

Main Methods:

  • Deep quantitative proteomics was performed on HUVEC cell lysates and conditioned medium.
  • HUVECs were stimulated with lipopolysaccharide (LPS) to activate TLR4.

Main Results:

  • Of 5205 detected proteins, 381 were classified as secreted.
  • 87.4% of secreted proteins used the conventional pathway, while 12.6% (48 proteins) utilized UCPS.
  • 43 UCPS-secreted proteins were newly identified, involved in redox regulation, proteostasis, and immune signaling.

Conclusions:

  • HUVEC secretome comprises both conventional and UCPS proteins regulating coagulation, angiogenesis, and immunity.
  • UCPS plays a significant role in endothelial cell secretion, with identified proteins like α-globin (HBA1) being crucial.
  • This study provides a valuable dataset for understanding UCPS mechanisms in endothelial cells.