Structural characterization of a diuretic peptide from the central nervous system of the leech Erpobdella octoculata. Angiotensin II Amide.
Purification of a material immunoreactive to an antiserum against angiotensin II and present in the central nervous system of the pharyngobdellid leech Erpobdella octoculata was performed by reversed-phase high pressure liquid chromatography combined with both enzyme-linked immunosorbent assay and dot immunobinding assays for angiotensin II. Establishment of the amino acid sequence by Edman degradation, electrospray, and fast atom bombardement mass spectrometry measurements and enzymatic treatment by carboxypeptidase A indicated that this "central" angiotensin II-like material, the first one fully characterized in the animal kingdom, is an angiotensin II amide. This finding constitutes also the first biochemical characterization of a peptide of the angiotensin family in an invertebrate. Synthetic angiotensin II amide exerts, when injected in leeches, a diuretic effect and is, 1 and 2 h postinjection, 100-fold more potent than vertebrate angiotensin II. An identification of the proteins immunoreactive to an antiserum against angiotensin II performed at the level of both central nervous system extracts and in vitro central nervous system-translated RNA products indicated that in the two cases, two proteins were detected. Their molecular masses, which were, respectively, approximately 14 and approximately 18 kDa for the central nervous system extracts and approximately 15 and approximately 19 kDa for in vitro central nervous system-translated RNA products, differ from that of angiotensinogen (approximately 60 kDa), the precursor of vertebrate angiotensin II.
Salzet
M
M
Laboratoire de Phylogénie moléculaire des Annélides ER 87 CNRS, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Bulet
P
P
Wattez
C
C
Verger-Bocquet
M
M
Malecha
J
J
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004232:Diuretics / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera; D015151:Immunoblotting; D007865:Leeches / Q000187:drug effects; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D009420:Nervous System / Q000737:chemistry*; D016339:Spectrometry, Mass, Fast Atom Bombardment
D004232:Diuretics; D007106:Immune Sera; D009419:Nerve Tissue Proteins; C091631:angiotensin II amide; D000804:Angiotensin II