• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. May 1, 2000; 347(Pt 3): 711–718.
PMCID: PMC1221007

Shedding of somatic angiotensin-converting enzyme (ACE) is inefficient compared with testis ACE despite cleavage at identical stalk sites.


The somatic and testis isoforms of angiotensin-converting enzyme (ACE) are both C-terminally anchored ectoproteins that are shed by an unidentified secretase. Although testis and somatic ACE both share the same stalk and membrane domains the latter was reported to be shed inefficiently compared with testis ACE, and this was ascribed to cleavage at an alternative site [Beldent, Michaud, Bonnefoy, Chauvet and Corvol (1995) J. Biol. Chem. 270, 28962-28969]. These differences constitute a useful model system of the regulation and substrate preferences of the ACE secretase, and hence we investigated this further. In transfected Chinese hamster ovary cells, human somatic ACE (hsACE) was indeed shed less efficiently than human testis ACE, and shedding of somatic ACE responded poorly to phorbol ester activation. However, using several analytical techniques, we found no evidence that the somatic ACE cleavage site differed from that characterized in testis ACE. First, anti-peptide antibodies raised to specific sequences on either side of the reported cleavage site (Arg(1137)/Leu(1138)) clearly recognized soluble porcine somatic ACE, indicating that cleavage was C-terminal to Arg(1137). Second, a competitive ELISA gave superimposable curves for porcine plasma ACE, secretase-cleaved porcine somatic ACE (eACE), and trypsin-cleaved ACE, suggesting similar C-terminal sequences. Third, mass-spectral analyses of digests of released soluble hsACE or of eACE enabled precise assignments of the C-termini, in each case to Arg(1203). These data indicated that soluble human and porcine somatic ACE, whether generated in vivo or in vitro, have C-termini consistent with cleavage at a single site, the Arg(1203)/Ser(1204) bond, identical with the Arg(627)/Ser(628) site in testis ACE. In conclusion, the inefficient release of somatic ACE is not due to cleavage at an alternative stalk site, but instead supports the hypothesis that the testis ACE ectodomain contains a motif that activates shedding, which is occluded by the additional domain found in somatic ACE.

Full Text

The Full Text of this article is available as a PDF (177K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ehlers MR, Riordan JF. Angiotensin-converting enzyme: new concepts concerning its biological role. Biochemistry. 1989 Jun 27;28(13):5311–5318. [PubMed]
  • Hooper NM. Angiotensin converting enzyme: implications from molecular biology for its physiological functions. Int J Biochem. 1991;23(7-8):641–647. [PubMed]
  • Hagaman JR, Moyer JS, Bachman ES, Sibony M, Magyar PL, Welch JE, Smithies O, Krege JH, O'Brien DA. Angiotensin-converting enzyme and male fertility. Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2552–2557. [PMC free article] [PubMed]
  • Ramaraj P, Kessler SP, Colmenares C, Sen GC. Selective restoration of male fertility in mice lacking angiotensin-converting enzymes by sperm-specific expression of the testicular isozyme. J Clin Invest. 1998 Jul 15;102(2):371–378. [PMC free article] [PubMed]
  • Soubrier F, Alhenc-Gelas F, Hubert C, Allegrini J, John M, Tregear G, Corvol P. Two putative active centers in human angiotensin I-converting enzyme revealed by molecular cloning. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9386–9390. [PMC free article] [PubMed]
  • Ehlers MR, Fox EA, Strydom DJ, Riordan JF. Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7741–7745. [PMC free article] [PubMed]
  • Howard TE, Shai SY, Langford KG, Martin BM, Bernstein KE. Transcription of testicular angiotensin-converting enzyme (ACE) is initiated within the 12th intron of the somatic ACE gene. Mol Cell Biol. 1990 Aug;10(8):4294–4302. [PMC free article] [PubMed]
  • Ehlers MR, Chen YN, Riordan JF. Spontaneous solubilization of membrane-bound human testis angiotensin-converting enzyme expressed in Chinese hamster ovary cells. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):1009–1013. [PMC free article] [PubMed]
  • Wei L, Alhenc-Gelas F, Soubrier F, Michaud A, Corvol P, Clauser E. Expression and characterization of recombinant human angiotensin I-converting enzyme. Evidence for a C-terminal transmembrane anchor and for a proteolytic processing of the secreted recombinant and plasma enzymes. J Biol Chem. 1991 Mar 25;266(9):5540–5546. [PubMed]
  • Oppong SY, Hooper NM. Characterization of a secretase activity which releases angiotensin-converting enzyme from the membrane. Biochem J. 1993 Jun 1;292(Pt 2):597–603. [PMC free article] [PubMed]
  • Ehlers MR, Riordan JF. Membrane proteins with soluble counterparts: role of proteolysis in the release of transmembrane proteins. Biochemistry. 1991 Oct 22;30(42):10065–10074. [PubMed]
  • Hooper NM, Karran EH, Turner AJ. Membrane protein secretases. Biochem J. 1997 Jan 15;321(Pt 2):265–279. [PMC free article] [PubMed]
  • Black RA, Rauch CT, Kozlosky CJ, Peschon JJ, Slack JL, Wolfson MF, Castner BJ, Stocking KL, Reddy P, Srinivasan S, et al. A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. Nature. 1997 Feb 20;385(6618):729–733. [PubMed]
  • Moss ML, Jin SL, Milla ME, Bickett DM, Burkhart W, Carter HL, Chen WJ, Clay WC, Didsbury JR, Hassler D, et al. Cloning of a disintegrin metalloproteinase that processes precursor tumour-necrosis factor-alpha. Nature. 1997 Feb 20;385(6618):733–736. [PubMed]
  • Parvathy S, Oppong SY, Karran EH, Buckle DR, Turner AJ, Hooper NM. Angiotensin-converting enzyme secretase is inhibited by zinc metalloprotease inhibitors and requires its substrate to be inserted in a lipid bilayer. Biochem J. 1997 Oct 1;327(Pt 1):37–43. [PMC free article] [PubMed]
  • Parvathy S, Karran EH, Turner AJ, Hooper NM. The secretases that cleave angiotensin converting enzyme and the amyloid precursor protein are distinct from tumour necrosis factor-alpha convertase. FEBS Lett. 1998 Jul 10;431(1):63–65. [PubMed]
  • Sadhukhan R, Santhamma KR, Reddy P, Peschon JJ, Black RA, Sen I. Unaltered cleavage and secretion of angiotensin-converting enzyme in tumor necrosis factor-alpha-converting enzyme-deficient mice. J Biol Chem. 1999 Apr 9;274(15):10511–10516. [PubMed]
  • Esther CR, Marino EM, Howard TE, Machaud A, Corvol P, Capecchi MR, Bernstein KE. The critical role of tissue angiotensin-converting enzyme as revealed by gene targeting in mice. J Clin Invest. 1997 May 15;99(10):2375–2385. [PMC free article] [PubMed]
  • Arribas J, Coodly L, Vollmer P, Kishimoto TK, Rose-John S, Massagué J. Diverse cell surface protein ectodomains are shed by a system sensitive to metalloprotease inhibitors. J Biol Chem. 1996 May 10;271(19):11376–11382. [PubMed]
  • Ramchandran R, Sen GC, Misono K, Sen I. Regulated cleavage-secretion of the membrane-bound angiotensin-converting enzyme. J Biol Chem. 1994 Jan 21;269(3):2125–2130. [PubMed]
  • Ehlers MR, Schwager SL, Scholle RR, Manji GA, Brandt WF, Riordan JF. Proteolytic release of membrane-bound angiotensin-converting enzyme: role of the juxtamembrane stalk sequence. Biochemistry. 1996 Jul 23;35(29):9549–9559. [PubMed]
  • Beldent V, Michaud A, Wei L, Chauvet MT, Corvol P. Proteolytic release of human angiotensin-converting enzyme. Localization of the cleavage site. J Biol Chem. 1993 Dec 15;268(35):26428–26434. [PubMed]
  • Beldent V, Michaud A, Bonnefoy C, Chauvet MT, Corvol P. Cell surface localization of proteolysis of human endothelial angiotensin I-converting enzyme. Effect of the amino-terminal domain in the solubilization process. J Biol Chem. 1995 Dec 1;270(48):28962–28969. [PubMed]
  • Schwager SL, Chubb AJ, Scholle RR, Brandt WF, Eckerskorn C, Sturrock ED, Ehlers MR. Phorbol ester-induced juxtamembrane cleavage of angiotensin-converting enzyme is not inhibited by a stalk containing intrachain disulfides. Biochemistry. 1998 Nov 3;37(44):15449–15456. [PubMed]
  • Schwager SL, Chubb AJ, Scholle RR, Brandt WF, Mentele R, Riordan JF, Sturrock ED, Ehlers MR. Modulation of juxtamembrane cleavage ("shedding") of angiotensin-converting enzyme by stalk glycosylation: evidence for an alternative shedding protease. Biochemistry. 1999 Aug 10;38(32):10388–10397. [PubMed]
  • Sadhukhan R, Sen GC, Ramchandran R, Sen I. The distal ectodomain of angiotensin-converting enzyme regulates its cleavage-secretion from the cell surface. Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):138–143. [PMC free article] [PubMed]
  • Wei L, Alhenc-Gelas F, Corvol P, Clauser E. The two homologous domains of human angiotensin I-converting enzyme are both catalytically active. J Biol Chem. 1991 May 15;266(14):9002–9008. [PubMed]
  • Ehlers MR, Chen YN, Riordan JF. Purification and characterization of recombinant human testis angiotensin-converting enzyme expressed in Chinese hamster ovary cells. Protein Expr Purif. 1991 Feb;2(1):1–9. [PubMed]
  • Hooper NM, Keen J, Pappin DJ, Turner AJ. Pig kidney angiotensin converting enzyme. Purification and characterization of amphipathic and hydrophilic forms of the enzyme establishes C-terminal anchorage to the plasma membrane. Biochem J. 1987 Oct 1;247(1):85–93. [PMC free article] [PubMed]
  • Baldwin SA. Use of antipeptide antibodies for the isolation and study of membrane proteins. Methods Mol Biol. 1994;27:43–63. [PubMed]
  • Williams TA, Barnes K, Kenny AJ, Turner AJ, Hooper NM. A comparison of the zinc contents and substrate specificities of the endothelial and testicular forms of porcine angiotensin converting enzyme and the preparation of isoenzyme-specific antisera. Biochem J. 1992 Dec 15;288(Pt 3):875–881. [PMC free article] [PubMed]
  • Lanzillo JJ, Fanburg BL. Development of competitive enzyme immunoassays for human serum angiotensin-1-converting enzyme: a comparison of four assay configurations. Anal Biochem. 1982 Oct;126(1):156–164. [PubMed]
  • Dasarathy Y, Stevens J, Lanzillo JJ, Fanburg BL. Elevation of angiotensin converting enzyme in bovine endothelial cells quantitated by an ELISA. Life Sci. 1990;47(10):883–889. [PubMed]
  • Hooper NM, Broomfield SJ, Turner AJ. Characterization of antibodies to the glycosyl-phosphatidylinositol membrane anchors of mammalian proteins. Biochem J. 1991 Jan 15;273(Pt 2):301–306. [PMC free article] [PubMed]
  • Ehlers MR, Scholle RR, Riordan JF. Proteolytic release of human angiotensin-converting enzyme expressed in Chinese hamster ovary cells is enhanced by phorbol ester. Biochem Biophys Res Commun. 1995 Jan 17;206(2):541–547. [PubMed]
  • Lanzillo JJ, Stevens J, Dasarathy Y, Yotsumoto H, Fanburg BL. Angiotensin-converting enzyme from human tissues. Physicochemical, catalytic, and immunological properties. J Biol Chem. 1985 Dec 5;260(28):14938–14944. [PubMed]
  • Rose-John S, Heinrich PC. Soluble receptors for cytokines and growth factors: generation and biological function. Biochem J. 1994 Jun 1;300(Pt 2):281–290. [PMC free article] [PubMed]
  • Esch FS, Keim PS, Beattie EC, Blacher RW, Culwell AR, Oltersdorf T, McClure D, Ward PJ. Cleavage of amyloid beta peptide during constitutive processing of its precursor. Science. 1990 Jun 1;248(4959):1122–1124. [PubMed]
  • Nishimura K, Yoshida N, Hiwada K, Ueda E, Kokubu T. Purification of angiotensin I-converting enzyme from human lung. Biochim Biophys Acta. 1977 Aug 11;483(2):398–408. [PubMed]
  • Ehlers MR, Chen YN, Riordan JF. The unique N-terminal sequence of testis angiotensin-converting enzyme is heavily O-glycosylated and unessential for activity or stability. Biochem Biophys Res Commun. 1992 Feb 28;183(1):199–205. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...