| Title: |
Quantitative Measurement of mRNA Levels by RT-PCR. |
| Authors: |
Walker, John M., Maguire, Janet J., Davenport, Anthony P., Lees, Delphine M., Khan, Noorafza Q., Barker, Stewart, Corder, Roger |
| Source: |
Peptide Research Protocols; 2002, p125-145, 21p |
| Abstract: |
The original description of endothelin-1 (ET-1) included the concept that a novel processing enzyme, referred to as endothelin-converting enzyme (ECE), was required for cleavage of the Trp21-Val22 bond in the biosynthetic intermediate big ET-1 (1). Initially it was thought that ECE could be a chymotrypsinlike enzyme because of the nature of the peptide bond being cleaved (1). The challenge of identifying ECE was soon taken up by many groups and a variety of different proteolytic activities were proposed as potential ECEs (2). By 1990, studies of endogenous ET-1 synthesis by cultured endothelial cells and investigations of the systemic pressor effect of intravenously administered big ET-1 indicated that the physiological ECE was a phosphoramidon-sensitive enzyme (3-5). Phosphoramidon was also shown to inhibit hydrolysis of exogenous big ET-1 by cultured vascular smooth muscle and endothelial cells (6). The pursuit of an ECE that could selectively hydrolyze big ET-1 and was inhibited by phosphoramidon led quickly to the purification and cloning of a metallopeptidase called endothelin-converting enzyme-1 (ECE-1) (7-9). Shortly after this, a second phosphoramidon-sensitive peptidase, with ∼59% structural homology to ECE-1, was cloned and called ECE-2 (10). ECE-1 and ECE-2 are members of a family of type II integral membrane peptidases that also includes neutral endopeptidase 24.11 and the KELL and PEX proteins (11). [ABSTRACT FROM AUTHOR] |
|
Copyright of Peptide Research Protocols is the property of Springer eBooks and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) |
| Database: |
Supplemental Index |
