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Protease activated receptor‐1 increases phosphorylation of endothelial nitric oxide synthase at site thr495 in human corpus cavernosum endothelial cells
Author(s) -
Mantey Jane Ama,
MotleyJohnson Evangeline D.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb445
Subject(s) - enos , phosphorylation , thrombin , nitric oxide synthase , nitric oxide , signal transduction , western blot , receptor , endothelial dysfunction , microbiology and biotechnology , blot , medicine , endothelium , nitric oxide synthase type iii , endocrinology , chemistry , biology , pharmacology , biochemistry , platelet , gene
Endothelial dysfunction, a common symptom of vascular diseases such as erectile dysfunction (ED), encompasses a broad array of physical and molecular defects of the endothelium that are due to low nitric oxide (NO) availability. Protease activated receptor‐1 (PAR1) induces downstream phosphorylation of endothelial nitric oxide synthase (eNOS) at site Threonine‐495 (Thr495) to inhibit NO. We hypothesize that excessive phosphorylation at this site reduces NO availability and contributes to ED, a risk factor for cardiovascular disease. Immortalized human corpus cavernosum endothelial cells (HCCECs) were lysed and PAR1 expression was examined by Western blot analysis. HCCECs were stimulated with either thrombin (10 U/mL) or TFLLR (10μM), the PAR1 activating peptide, at varying time points and eNOS‐Thr495 phosphorylation was determined by Western blot. Results were quantified by densitometry, and demonstrated that PAR1 is expressed in HCCECs. Treatment with thrombin and TFLLR induced maximal downstream phosphorylation of eNOS‐Thr495 at 10 minutes. We believe that this signal transduction pathway is amplified in males with ED, and inhibition of PAR1 will restore NO availability. Our goal is to contribute to the development of novel therapeutic agents that focus on improving NO availability in ED and other cardiovascular diseases. This project was supported, in part, by NIGMS Grant 5SC3GM086336‐02.