Honors Theses

Date of Award

2008

Document Type

Undergraduate Thesis

Department

Chemistry and Biochemistry

First Advisor

Ziaeddin Shariat-Madar

Relational Format

Dissertation/Thesis

Abstract

Prolylcarboxypeptidase (PRCP) is involved in the conversion of angiotensin II (Ang II, a potent vasoconstrictor) to angiotensin 1-7 and conversion of bradykinin (BK, a vasodilator) to form des Arg9 bradykinin. PRCP converts plasma prekallikrein (PK) to plasma kallikrein when the high molecular weight kininogen (HK) combines with PK, forming the HK-PK complex that binds to human umbilical vein endothelial cells (HUVEC) membranes. Formed kallikrein then liberates BK jfrom HK, which leads to vasodilation. This physiological function is mediated by the BK type 2 receptor in the G protein-coupled receptor family. The balance between BK production and Ang II inactivation is important for wound healing, angiogenesis and high blood pressure regulation. Our goal was to determine the post translational regulation of PRCP in Chinese hamster ovary (CHO) cells. CHO cells were transfected with full-length PRCP under the control of a cytomegalovirus (CMV) promoter and rPRCP was expressed as a fusion protein with C-terminal enhanced green fluorescent protein (EGFP). The binding of biotinylated HK to wild type CHO cells was time dependent, dose dependent. saturable and reversible. The PRCP-induced PK activation was similar on wild type and PRCP-transfected CHO cells. PRCP inhibitor, Z-Pro-Prolinal did not block PK activation on wild type or PRCP-transfected CHO cells. In addition, since PRCP-induced PK activation was similar on both wild type and PRCP-transfected CHO cells; there is a possibility of the existence of a novel enzymatic substance that activates the kallikreinkinin system (KKS).

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