Determining the Enzymatic Activity of Angiotensin-Converting Enzyme 2 (ACE2) in Brain Tissue and Cerebrospinal Fluid Using a Quenched Fluorescent Substrate

Methods Mol Biol. 2017:1527:117-126. doi: 10.1007/978-1-4939-6625-7_9.

Abstract

Angiotensin-converting enzyme 2 (ACE2) is a component of the renin-angiotensin system (RAS) which plays an important role in the regulation of blood pressure and volume homeostasis. Accumulating evidence shows alterations in ACE2 expression and activity in several hypertensive animal models, as well as in patients with hypertension. In order to assess the role of brain ACE2 in hypertension, a specific ACE2 assay is required. Based on a quenched fluorescent substrate, we describe an easy-to-use method for determining ACE2 activity in brain tissue and cerebrospinal fluid. The method can further be adapted for other tissues, plasma, cell extracts, and cell culture supernatants.

Keywords: ACE2; Brain ACE2; CSF; DX600; Hypertension; Mca-APK(Dnp); Quenched fluorescent substrate.

MeSH terms

  • Angiotensin-Converting Enzyme 2
  • Animals
  • Blood Pressure / genetics
  • Blood Pressure / physiology
  • Brain / metabolism*
  • Enzyme Assays / methods*
  • Humans
  • Hypertension / cerebrospinal fluid
  • Hypertension / metabolism
  • Peptides
  • Peptidyl-Dipeptidase A / cerebrospinal fluid
  • Peptidyl-Dipeptidase A / metabolism*
  • Renin-Angiotensin System / physiology

Substances

  • DX600 peptide
  • Peptides
  • Peptidyl-Dipeptidase A
  • ACE2 protein, human
  • Angiotensin-Converting Enzyme 2