Quantitative Measurement of mRNA Levels by RT-PCR

Studies of ECE-1 Isoforms
  • Delphine M. Lees
  • Noorafza Q. Khan
  • Stewart Barker
  • Roger Corder
Part of the Methods in Molecular Biology™ book series (MIMB, volume 206)


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, 4, 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, 8, 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).


Pulmonary Artery Smooth Muscle Cell Amplification Plot Repeated Pipeting Dissociation Protocol Molecular Analyst Software 
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Copyright information

© Humana Press Inc. 2002

Authors and Affiliations

  • Delphine M. Lees
    • 1
  • Noorafza Q. Khan
    • 1
  • Stewart Barker
    • 1
  • Roger Corder
    • 1
  1. 1.Department of Experimental Therapeutics, William Harvey Research Institute, St. Bartholomew’es and the Royal London School of Medicine and DentistryQueen Mary University of LondonLondonUK

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