Functional residues at the active site of angiotensin converting enzyme

Bünning, Peter; Holmquist, Barton; Riordan, James

doi:10.1016/0006-291x(78)91382-7

Cited by 82 publications

(29 citation statements)

References 19 publications

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“…Our preliminary data suggest that at least two of the residues previously identified as important active-site residues, a tyrosine and a lysine (29,30), are located only in the C-terminal or "testicular" half of lung ACE (Y. N. Chen and J.F.R., unpublished results), confirming the hypothesis that the testis ACE active site is also the functional site in lung ACE. Elucidating the catalytic mechanism of ACE in molecular terms could thus most profitably be accomplished with the testis isozyme, since it is unencumbered by the (apparently) catalytically unimportant N-terminal domain of lung ACE.…”

Section: Discussionsupporting

confidence: 65%

Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

Ehlers

Fox

Strydom

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

281

161

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Angiotensin-converting enzyme (ACE; EC 3.4.15.1) is a zinc-containing dipeptidyl carboxypeptidase widely distributed in mammalian tissues and is thought to play a critical role in blood pressure regulation. Testis contains a unique, androgen-dependent ACE isozyme of unknown function. We have determined the cDNA sequence for human testicular ACE; it encodes a protein that is identical, from residue 37 to its C terminus, to the second half or C-terminal domain of the endothelial ACE sequence [Soubrier, F., AlhencGelas, F., Hubert, C., Allegrini, J., John, M., Tregear, G. & Corvol, P. (1988) Proc. Nati. Acad. Sci. USA 85, 9386-9390]. The full-length human testis ACE cDNA was constructed from a composite of cloned cDNAs, obtained by a combination of (i) immunoscreening and hybridization screening of a human testicular cDNA library in Agtll and (ii) hybridization screening of human testis cDNAs constructed with ACE-specific primers and amplified by the polymerase chain reaction. The protein sequence inferred consists of a 732-residue preprotein including a 31-residue signal peptide. The mature polypeptide has a molecular weight of 80,073. The testis enzyme contains the second of the two putative metal-binding sites (HisGlu-Met-Gly-His) identified in endothelial ACE. This indicates that the functionally active catalytic site is within the Cterminal domain of the endothelial enzyme, accounting for the previous finding that these two structurally dissimilar isozymes are virtually identical catalytically. Of 22 testis ACE cDNAs cloned and sequenced, 3 have unique 5' regions, consisting of inserted, deleted, or substituted sequences up to 328 base pairs long, which have apparently arisen by alternative pre-mRNA splicing.

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Section: Discussionsupporting

confidence: 65%

Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

Ehlers

Fox

Strydom

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

281

161

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“…This acetylation has been described as a rather unlikely possibility (Riordan and Vallee, 1972 b;Roskoski, 1974). The observed inactivation at pH 8.1 and its stability below pH 7.2 strongly argue against the acetylation of a histidine in PGH synthase (Riordan and Vallee, 1972a;Roskoski, 1974;Biinning et al, 1978). Furthermore, to our knowledge, there is no example for the acetylation by N-acetylimidazole of histidine in a protein documented in the literature.…”

Section: Discussionmentioning

confidence: 81%

“…1971 ;Riordan and Vallee, 1972a;Masiak and D'Angelo, 1975;Biinning et al, 1978) where spontaneous reactivation of PGH synthase was observed. The observed reactivation with hydroxylamine at pH 7.2 corroborates the exclusion of serine, threonine and lysine, which cannot be deacetylated with hydroxylamine below pH 10 (Balls and Wood, 1956;Riordan and Vallee, 1972a).…”

Section: Discussionmentioning

confidence: 99%

Chemical modification of prostaglandin endoperoxide synthase by N‐acetylimidazol

Karthein

Strieder

Ruf

1992

European Journal of Biochemistry

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Prostaglandin H synthase apoprotein, without its prosthetic heme group, was inactivated by Nacetylimidazole under conditions typical for the O-acetylation of tyrosyl residues. A spontaneous reactivation occurred above pH 7.5 at 22 "C, which indicated spontaneous hydrolysis of acetylated residues. Below pH 7.5, where stable inactivation was observed, reactivation was achieved by reaction with hydroxylamine. Both enzymic activities of prostaglandin H synthase, cyclooxygenase and peroxidase, were inactivated and reactivated simultaneously and to the same extent. In contrast to the apoprotein, the holoenzyme with heme was not inactivated by N-acetylimidazole. The number of acetyl groups, as determined as hydroxamate after the reaction with hydroxylamine at pH 8.2, was 2.5 k 0.4 for the apoprotein and 1 .O If: 0.24 for the holoenzyme. The specific binding of heme as the prosthetic group was no longer observed by EPR (signals at g = 6.7 and 5.3) when hemin was added to the N-acetylimidazole-reacted apoprotein. Treatment of N-acetylimidazole-reacted apoprotein with hydroxykamine restored the specific binding of heme. The N-acetylimidazole-reacted apoprotein supplemented with hemin and reacted with hydroperoxides, neither showed electronic absorption spectra of higher oxidation states nor an EPR doublet signal due to a tyrosyl radical.These results demonstrate that heme protects against the inactivating modification by Nacetylimidazole and that this modification prevents binding of the prosthetic heme group necessary for both enzymic activities. The absence of the prosthetic heme group explains the concomitant loss of cyclooxygenase and peroxidase activities, as well as the absence of higher oxidation states and the tyrosyl radical. We suggest that the acetylation of a residue in the heme pocket, most probably a tyrosine, although a histidine cannot be definitely disproved, exerts the inhibiting effect. This residue could be the axial ligand of the heme or in close contact to the heme. The results also show that the inhibition by N-acetylimidazole does not involve the acetylation of Ser530 which causes the inhibition by acetylsalicylic acid of cyclooxygenase. [The numbering of amino acids in ovine prostaglandin H synthase is according to DeWitt, D.

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“…In addition, most inhibitors of the two peptidases incorporate essentially similar featuresi.e., a zinc-chelating group borne by a modified dipeptide with a free C-terminal carboxylate interacting with a positively charged, presumably guanidinium residue (30,31), identified as Arg95 in rabbit enkephalinase (32). However, the optimal dipeptide sequences binding to the S; and S' subsites (nomenclature according to ref.…”

Section: Resultsmentioning

confidence: 99%

Mixed inhibitors of angiotensin-converting enzyme (EC 3.4.15.1) and enkephalinase (EC 3.4.24.11): rational design, properties, and potential cardiovascular applications of glycopril and alatriopril.

Gros

Nadine

Souque

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

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Two major hormonal peptide systems appear to play opposite roles in the regulation of electrolyte balance and blood pressure, so that their imbalance might be responsible for cardiovascular and salt-retention disorders affecting a large fraction of the population (1). The first one is the reninangiotensin-aldosterone system, whose critical role in sodium retention and vasoconstriction was largely evidenced through the use of inhibitors of the membra~ne metallopeptidase angiotensin-converting enzyme (ACE; EC 3.4.15.1) (2, 3). These drugs, which prevent the formation of angiotensin II, a vasoactive, antidiuretic, and antinatriuretic peptide, and decrease blood pressure and aldosterone secretion, have gained wide application in the treatment of essential hypertension and congestive heart failure. The second hormonal system is constituted by the atrial natriuretic factor (ANF), a peptide secreted by the heart into the circulation to decrease blood pressure, raise the urinary excretion of water and sodium, and lower plasma renin and aldosterone levels (4). Recently, the role of another metallopeptidase, enkephalinase (EC 3.4.24.11, membrane metalloendopeptidase), in the inactivation of endogenous ANF was shown by the effects of inhibitors (reviewed in ref. 5). These drugs enhance the circulating level of the hormone in healthy volunteers as well as in patients with congestive heart failure or cirrhosis; induce diuresis, natriuresis, and urinary excretion of cGMP (6-10); and may exert antihypertensive activity (11).Hence it appears that two cell surface, zinc-containing peptidases play a key role in the activation or inactivation pathways of the two hormonal systems. Although many compounds are highly selective inhibitors of either ACE or enkephalinase, some mercaptoalkyl inhibitors display limited but significant crossreactivity toward the two peptidases (12,13). This partial overlap suggested that a careful analysis of structure-activity relationships would enable the design of a single molecular structure able to potently inhibit both enzymes in vivo and, thereby, lead to a class of potentially useful cardiovascular agents blocking both the generation of angiotensin II and the inactivation of ANF.We have now designed such mixed inhibitors which interact with the two peptidases at nanomolar concentrations in vitro and at low dosage per os. These drugs elicit the characteristic actions ofthe two classes ofenzyme inhibitors: prevention of angiotensin I-induced hypertension, protection of ANF, enhancement of diuresis and natriuresis, and increase in urinary cGMP excretion.

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Functional residues at the active site of angiotensin converting enzyme

Cited by 82 publications

References 19 publications

Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

Chemical modification of prostaglandin endoperoxide synthase by N‐acetylimidazol

Mixed inhibitors of angiotensin-converting enzyme (EC 3.4.15.1) and enkephalinase (EC 3.4.24.11): rational design, properties, and potential cardiovascular applications of glycopril and alatriopril.

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