Human Transmembrane Serine Protease 2 (TMPRSS2) on Human Seminal Fluid Extracellular Vesicles Is Proteolytically...

Area of Science:

• Biochemistry
• Molecular Biology
• Virology

Background:

• Human transmembrane serine protease 2 (TMPRSS2) is crucial for viral entry and disease pathogenesis.
• Targeting TMPRSS2 requires a stable, active enzyme source for research and drug development.

Purpose of the Study:

• To characterize native, active TMPRSS2 on seminal fluid extracellular vesicles (SF-EVs).
• To develop a robust in vitro assay for screening TMPRSS2 inhibitors.
• To compare SF-EV-associated TMPRSS2 with recombinant forms.

Main Methods:

• Characterization of SF-EVs bearing surface-exposed TMPRSS2 using flow cytometry and activity assays.
• Recombinant production of human TMPRSS2 ectodomain.
• Biochemical assays to determine catalytic parameters and inhibition profiles.
• Development of an SF-EV-based in vitro screening assay.

Main Results:

• SF-EVs present enzymatically active, surface-exposed TMPRSS2.
• Recombinant TMPRSS2 ectodomain was successfully produced.
• Native SF-EV-associated and recombinant TMPRSS2 showed comparable activity and inhibition by Nafamostat mesylate.
• A robust SF-EV-based assay for TMPRSS2 inhibitor screening was established.

Conclusions:

• SF-EVs provide a stable, native source of active TMPRSS2.
• The developed assay facilitates the discovery of TMPRSS2-targeting therapeutics.
• Further research into SF-EV-associated TMPRSS2's biological roles is warranted.