Could angiotensin I be produced from a renin substrate by the HIV-1 protease?
The standard angiotensin I (Ang I) radioimmunoassay for renin activity determination is a useful clinical tool for the diagnosis of high renin levels in certain cases of hypertension. It depends upon the liberation of Ang I from human plasma angiotensinogen. We considered whether a commercially available synthetic tetradecapeptide (TDP), Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val-Tyr-Ser, would produce authentic Ang I upon incubation with protease from human immunodeficiency virus type 1 (HIV-1). This peptide is also known to be cleaved by renin at the Leu-Leu bond to yield the decapeptide Ang I. When the TDP is incubated with the HIV-1 protease, the peptide is readily hydrolyzed. Product formation is linear with respect to time and enzyme concentration. HPLC analysis of reaction products showed two new peaks, as one would expect from the cleavage of a TDP into a decapeptide and a tetrapeptide. Amino acid analysis of HPLC-purified peaks confirmed that the HIV-1 protease cleaves TDP at the Leu10-Leu11 site to produce the desired decapeptide, Ang I. Production of Ang I by the HIV-1 protease, like human renin, is inhibited in the presence of a protease inhibitor. Implications of the discovery of an HIV-1 protease substrate that produces authentic Ang I are discussed in light of a screening assay for soluble HIV-1 protease inhibitors.
Sharma
S K
SK
Department of Biochemistry, Upjohn Laboratories, Upjohn Company, Kalamazoo, Michigan 49001.
Evans
D B
DB
Hui
J O
JO
Heinrikson
R L
RL
D000595:Amino Acid Sequence; D000803:Angiotensin I / Q000096:biosynthesis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D016333:HIV Protease / Q000737:chemistry*; D017320:HIV Protease Inhibitors; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011480:Protease Inhibitors / Q000494:pharmacology; D012083:Renin / Q000737:chemistry*; D012995:Solubility; D013379:Substrate Specificity
D017320:HIV Protease Inhibitors; D010455:Peptides; D011480:Protease Inhibitors; D000803:Angiotensin I; D016333:HIV Protease; D012083:Renin