Isolation of a renin-like enzyme from the leech Theromyzon tessulatum.
This article reports the purification of a renin-like enzyme (an aspartyl protease) from head parts of the leech Theromyzon tessulatum. After four steps of purification including gel permeation and anion exchange chromatographies followed by reversed-phase HPLC, this enzyme was purified to homogeneity. The renin-like enzyme (of 32 kDa) hydrolyses at neutral pH and at 37 degrees C, the Leu10-Leu11 bond of synthetic porcine angiotensinogen tetradecapeptide yielding the angiotensin I and the Leu11-Val12-Tyr13-Ser14 peptide as products, with a specific activity of 1.35 pmol AI/min/mg (Km 22 microM; Kcat 2.7). The hydrolysis of angiotensinogen is inhibitable at 90% by pepstatin A (IC50 = 4.6 microM), consistent with a renin activity. This is the first biochemical evidence of renin-like enzyme in invertebrates.
Laurent
V
V
Laboratoire de Phylogénie moléculaire des Annélides, ER 87 CNRS, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Salzet
M
M
D000595:Amino Acid Sequence; D000803:Angiotensin I / Q000235:genetics / Q000378:metabolism; D000804:Angiotensin II / Q000235:genetics / Q000378:metabolism; D000808:Angiotensinogen / Q000235:genetics / Q000378:metabolism; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001665:Binding Sites; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007865:Leeches / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012083:Renin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity; D013552:Swine; D013696:Temperature
D010455:Peptides; D000808:Angiotensinogen; D000804:Angiotensin II; D000803:Angiotensin I; D016282:Aspartic Acid Endopeptidases; D012083:Renin