TMPRSS2 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

TMPRSS2, also called Epitheliasin in mice, is a 492 amino acid type II transmembrane serine protease located on human chromosome 21q22.3 that encodes at least 2 isoforms. This glycosylated serine protease (theoretical molecular weight 70kDa) is regulated by androgens and expressed on the plasma membrane of human bronchial epithelial cells, nasal goblet cells, small intestine epithelia, gastrointestinal tract, the stomach, kidneys and pancreas, with the greatest abundance found in the prostate gland. While the physiological function of TMPRSS2 remains unknown, the serine protease domain undergoes autocleavage and the 32 kDa domain is secreted enabling its interaction with the extracellular matrix. Fusions between TMPRSS2 and the ETS transcription factor genes ERG, ETV1, and ETV4 have been reported in prostate cancer with the TMPRSS2 ERG gene fusion resulting in ERG overexpression in 40-80% of these cases and often producing a more aggressive phenotype. Androgen signaling is disrupted in prostate cancers with the TMPRSS2 ERG fusion which contributes to the switch from androgen dependent to androgen independent prostate cancer (1,2).

TMPRSS2 has also been shown to play a critical role in the process of viral entry for coronaviruses and influenza viruses thru proteolytic activation of key viral glycoproteins. One pathway for SARS-CoV-2 infection, which causes the COVID-19 disease, involves binding of the SARS-CoV-2 Spike 'S' glycoprotein to the human ACE2 receptor and cleavage of S by TMPRSS2 at the cell surface to facilitate viral entry. TMPRSS2 mediates viral entry in a similar mechanism for other coronaviruses such as SARS-CoV and MERS. The broad-spectrum serine protease inhibitor, Camostat, is a TMPRSS2 inhibitor demonstrated to protect mice with lethal SARS-CoV infections (3).

References

1. Clark, J., Cooper, C. (2009) ETS gene fusions in prostate cancer. Nat Rev Urol 6, 429-439. PMID: 19657377

2. Duffy, MJ. (2014) Chapter One - PSA in Screening for Prostate Cancer: More Good than Harm or More Harm than Good? Adv Clin Chem. 66:1-23. PMID: 25344984

3. Shen LW, Mao HJ, Wu YL, Tanaka Y, Zhang W. (2017) TMPRSS2: A potential target for treatment of influenza virus and coronavirus infections. Biochimie. 142:1-10

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBL1-17121

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• TMPRSS2 ELISA Kits
• TMPRSS2 Lysate
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