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Student Contribution  |   January 2005
Demonstration of Endopeptidase 24.15 in Rabbit Heart: Evidence of Bradykinin's Role in Cardioprotection
Article Information
Cardiovascular Disorders
Student Contribution   |   January 2005
Demonstration of Endopeptidase 24.15 in Rabbit Heart: Evidence of Bradykinin's Role in Cardioprotection
The Journal of the American Osteopathic Association, January 2005, Vol. 105, 21. doi:10.7556/jaoa.2005.105.1.21
The Journal of the American Osteopathic Association, January 2005, Vol. 105, 21. doi:10.7556/jaoa.2005.105.1.21
Web of Science® Times Cited: 10
Bradykinin (BK) plays an important role as an endogenous mediator resulting in acute protection from myocardial ischemia reperfusion injury. Shielding BK from metabolism prolongs its activity and increases protection during reperfusion following an ischemic event. EC 3.4.24.15 (EP24.15) is one of the enzymes responsible for the metabolism of BK. Previous work in this lab has shown inhibitors of EP24.15 increased BK levels in the heart and decreased reperfusion injury, but the actual enzyme's presence has not been validated in heart tissue. The goal of this study was to demonstrate that EP24.15 is present in the heart. 
Partially purified rabbit heart homogenates were separated by 10% SDS–12% polyacrylamide gel electrophoresis and underwent Western Blot analysis. The nitrocellulose membranes were incubated with anti-EP24.15 antibody (1:1500 dilution) overnight. The secondary antibody (goat anti–mouse IgG HRP in a 1:2000 dilution) was incubated for 1 hour at room temperature. The membranes underwent chemiluminescence and were exposed to x-ray film. 
The analysis showed bands of protein which correspond to authentic EP24.15 in the heart tissue samples. These results demonstrate EP24.15's presence in rabbit heart tissue, thus providing support for the idea that inhibitors of this enzyme work by inhibiting the actual EP24.15 enzyme and not by other nonspecific mechanisms.