Eur. J. Entomol. 100 (4): 467-474, 2003 | DOI: 10.14411/eje.2003.070

Isolation of angiotensin converting enzyme from testes of Locusta migratoria (Orthoptera)

Nathalie MACOURS1,*, Anick VANDINGENEN1, Constant GIELENS2, Korneel HENS1, Geert BAGGERMAN1, Liliane SCHOOFS1, Roger HUYBRECHTS1
1 Laboratory of Developmental Physiology and Molecular Biology, Katholieke Universiteit Leuven, Naamsestraat 59, B-3000 Leuven, Belgium
2 Laboratory of Biochemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200G, B-3001 Leuven-Heverlee, Belgium

By means of a tracer assay using a labeled synthetic angiotensin converting enzyme (ACE) substrate hippurylglycylglycine, we have detected high ACE activity in the testes of the African migratory locust, Locusta migratoria. Lower, but significant, ACE activity was observed in midgut and hemolymph. In a two-step purification procedure involving anion exchange and gel permeation chromatography, we have purified LomACE from the locust testes. The enzyme of approximately 80 kDa shows substantial amino-acid sequence homology with ACE from both vertebrate and invertebrate origin. The ACE identity of the purified enzyme was further confirmed by cDNA cloning of the Locusta ACE fragment, which, after in silico translation, revealed a mature protein of 623 amino acids with a large structural similarity to other known ACE proteins.

Keywords: ACE inhibitors, cDNA cloning, insects, Locusta migratoria, reproduction, testes

Received: January 13, 2003; Accepted: June 26, 2003; Published: November 20, 2003

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