Renin assay using a fluorogenic substrate and high performance liquid chromatography.
Measurement of renin activity in human fluids using a fluorogenic substrate and high performance liquid chromatography (HPLC) is described. A nine amino acid peptide containing the fluorogenic residue, N-(2-pyridyl) glycine (Pg) is used as a substrate. The peptide sequence is homologous with the cleavage site of human angiotensinogen. This substrate is hydrolyzed by renin to generate fluorogenic and non-fluorogenic products. The amount of fluorogenic product is directly measured by reversed phase HPLC. Optimization of assay conditions and measurement of human serum renin levels are described. Assay results correlated well with those from radioimmunoassay. The method is simple, convenient, highly sensitive and can be used for routine clinical renin assays.
Nakamura-Imajo
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N
Osaka Research Laboratory, Wako Pure Chemical Industries, Ltd., Hyogo, Japan.
Satomura
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S
Matsuura
S
S
Murakami
K
K
D000595:Amino Acid Sequence; D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D010450:Endopeptidases / Q000097:blood; D004792:Enzyme Precursors / Q000097:blood; D005456:Fluorescent Dyes; D005998:Glycine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011725:Pyridines / Q000737:chemistry / Q000378:metabolism*; D011786:Quality Control; D012016:Reference Values; D012083:Renin / Q000097:blood*
D002021:Buffers; D004792:Enzyme Precursors; D005456:Fluorescent Dyes; D010446:Peptide Fragments; D011725:Pyridines; C071078:N-(2-pyridyl)glycine; D010450:Endopeptidases; D012083:Renin; D005998:Glycine