Abstract
Angiotensin-converting enzyme (ACE) is a dipeptidyl carboxypeptidase that converts angiotensin I into the potent vasoconstrictor angiotensin II. We have used cDNA and genomic sequences to assemble a composite cDNA, ACE.315, encoding the entire amino acid sequence of mouse converting enzyme. ACE.315 contains 4838 base pairs and encodes a protein of 1278 amino acids (147.4 kDa) after removal of a 34-amino acid signal peptide. Within the protein, there are two large areas of homologous sequence, each containing a potential Zn-binding region and catalytic site. These homologous regions are approximately half the size of the whole ACE protein and suggest that the modern ACE gene is the duplicated product of a precursor gene. Mouse ACE is 83% homologous to human ACE in both nucleic acid and amino acid sequence, and like human ACE, contains a hydrophobic region in the carboxyl terminus that probably anchors the enzyme to the cell membrane (Soubrier, F., Alhenc-Gelas, F., Hubert, C., Allegrini, J., John, M., Tregear, G., and Corvol, P. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 9386-9390). Northern analysis of mouse kidney, lung, and testis RNA demonstrates that the testicular isozyme of ACE is encoded by a single, smaller RNA (2500 bases) than the two message sizes found in kidney or lung (4900 and 4150 bases), and that this testicular RNA hybridizes to the 3' portion of ACE.315.
Highlights
Angiotensin-converting enzyme(ACE) is a dipepti- tation that its inhibition would lower blood pressure in padyl carboxypeptidase that converts angiotensin I into tients with high renin levels
We previously reported the sequence of ACE.31, stood, and such basic questions as its amino acid structure a cDNA that encodes the NHz-terminalamino acid sequence are only being addressed
3’ these two cDNA are completely divergent with ACE.5 encoding a total of 873nucleotides of 3’ Ut and ACE.ll encoding 1751 nucleotides of 3’ Ut (Fig. 2)
Summary
MRNA was prepared from the kidneys of male NIH Swiss mice and selected by oligo(dT)chromatography [17]. The construction of a mouse kidney cDNA library, the screening of this library with ACE-specific oligonucleotide probes, the subcloning of double-stranded cDNA into the EcoRI site of Bluescript (+)(Stratagene, San Diego,CA), and the rescue and sequencing of single-stranded DNA have been described [15].During the construction of cDNA, internal EcoRI sites were not protected by methylation resulting in the separate cloning of sequences on either side of internal EcoRI sites (GAATTC). ACE.ll was completely sequenced on both strands using a combination of three approaches, subcloning RsaI restriction fragments of ACE.ll into theSmoI site of mplO M13,priming single-stranded DNA with complementary ACE oligonucleotides and unidirectional progressive deletions using EzoIII nuclease A 3200bp PstI restriction fragment was found to hybridize with both ACE. and ACE.ll probes Both ACE.31/557 and ACE.11/ 514 were isolated free of extraneous DNA by agarose gel electropho-
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
