Cloning and Expression of an Evolutionary Conserved Single-domain Angiotensin Converting Enzyme from Drosophila melanogaster(*)
- Michael J. Cornell(1),
- Tracy A. Williams(3),
- Nazarius S. Lamango(1),
- David Coates(1),
- Pierre Corvol(3),
- Florent Soubrier(3),
- Jorg Hoheisel(4),
- Hans Lehrach(2) and
- R. Elwyn Isaac(1)(§)
- From the (1) Department of Pure & Applied Biology, University of Leeds, Leeds LS2 9JT, United Kingdom, and the
- (2) Genome Analysis Laboratory, Imperial Cancer Research Fund, Lincoln Inn's Fields, London WC2A 3PX, United Kingdom,
- (3) INSERM Unit 36, Laboratoire de Medecine Experimentale, College de France, 3 rue de Ulm, 75005 Paris, France, and the
- (4) Molecular-Genetic Genome Analysis Laboratory, Deutsches Krebsforschungszentrum im Neuenheimer Feld 280, D-69120 Heidelberg, Federal Republic of Germany
- § To whom correspondence should be addressed: Dept. of Pure & Applied Biology, University of Leeds, Leeds LS2 9JT, United Kingdom. Tel.: 44-113-233-2903; Fax: 44-113-233-2882.
Abstract
Mammalian somatic angiotensin converting enzyme (EC 3.4.15.1, ACE) consists of two highly homologous (N- and C-) domains encoded
by a duplicated gene. We have identified an apparent single-domain (67 kDa) insect angiotensin converting enzyme (AnCE) in
embryos of Drosophila melanogaster which converts angiotensin I to angiotensin II (K
, 365 μM), removes Phe-Arg from the C terminus of bradykinin (K
, 22 μM), and is inhibited by ACE inhibitors, captopril (IC
= 1.1 
10
M) and trandolaprilat (IC
= 1.6 
10
M). We also report the cloning and expression of a Drosophila AnCE cDNA which codes for a single-domain 615-amino acid protein with a predicted 17-amino acid signal peptide and regions
with high levels of homology to both the N- and C-domains of mammalian somatic ACE, especially around the active site consensus
sequence. Northern analysis identified a single 2.1-kilobase mRNA in Drosophila embryos, and Southern analysis of Drosophila genomic DNA indicates that the insect gene is not duplicated. When expressed in COS-7 cells, the AnCE protein is a secreted
enzyme, which converts angiotensin I to angiotensin II and is inhibited by captopril (IC
= 5.6 
10
M) and trandolaprilat (IC
= 2 
10
M). The evolutionary significance of these results is discussed.
Footnotes
-
↵* This work was supported by Grant GR/F96792 from the Science and Engineering Research Council, United Kingdom, by grants from the Fondation pour la Recherche Medicale, France and by a Science and Engineering Research Council research studentship (to M. J. C.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank
/EMBL Data Bank with accession number(s) U25344.
-
↵1 The abbreviations used are:
- ACE
-
angiotensin converting enzyme
- sACE
-
somatic ACE
- tACE
-
testicular ACE
- AnCE
-
D. melanogaster ACE
- AI
-
angiotensin I
- AII
-
angiotensin II
- BK
-
bradykinin
- bp
-
base pairs
- CHAPS
-
3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate
- HPLC
-
high performance liquid chromatography
- kb
-
kilobase pairs
- PAGE
-
polyacrylamide gel electrophoresis.
-
↵2N. S. Lamango and R. E. Isaac, manuscript in preparation.
- © 1995 by The American Society for Biochemistry and Molecular Biology, Inc.
