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Bimane fluorogenic substrates for microdetermination of angiotensin converting enzyme level in serum
E Sato1, H Hattori, S Nishikawa
1Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Chemical & Pharmaceutical Bulletin
|August 1, 1991
Summary
This study shows that bimane peptides with nitrated amino acids are effective fluorogenic substrates for measuring angiotensin I converting enzyme (ACE). Substrate 3 is particularly potent for determining ACE levels in human serum.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Enzyme Assays
Background:
- 9,10-dioxa-syn-3,4,6,7-tetramethylbimane (bimane) fluorescence is quenched by nitrated aromatic amino acids.
- Bimane peptides offer potential as fluorogenic substrates for enzyme activity determination.
Purpose of the Study:
- To develop and evaluate bimane peptides containing nitrated amino acids as fluorogenic substrates for angiotensin I converting enzyme (ACE) assays.
- To compare the efficacy of these novel substrates with previously reported bimane substrates.
Main Methods:
- Synthesis of bimane peptides incorporating nitrated amino acids.
- Enzyme kinetic assays using rabbit lung ACE.
- Fluorogenic detection of enzyme activity.
- Microdetermination of ACE levels in human serum samples.
Main Results:
- Bimane fluorescence quenching was observed with nitrated aromatic amino acids.
- Bimane peptides with nitrated amino acids (1a, b) demonstrated utility as fluorogenic substrates for rabbit lung ACE.
- Substrate 3, a bimane peptide containing tryptophan, proved to be a potent fluorogenic substrate for ACE.
- Substrate 3 enabled the microdetermination of ACE levels in human serum.
Conclusions:
- Nitrated amino acid-containing bimane peptides are effective fluorogenic substrates for ACE.
- Substrate 3 is a highly sensitive and potent tool for quantifying ACE in human serum, facilitating microanalysis.