Purification of Angiotensin-Converting Enzyme from Human Intestine

Hayakari, Makoto; Amano, Kayo; Izumi, Hiroshi; Murakami, Satoshi

doi:10.1007/978-1-4615-9546-5_60

Cited by 3 publications

(2 citation statements)

References 15 publications

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“…Another possibility for biogenesis of ileal ACE would involve alternate splicing of its mRNA, yielding a single N-domain enzyme directly, although ACE purified from human and rat intestinal tissue is the full-size enzyme (43)(44)(45).…”

Section: Resultsmentioning

confidence: 99%

Naturally occurring active N-domain of human angiotensin I-converting enzyme.

Deddish

Wang

Michel

et al. 1994

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

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Angiotensin I-converting enzyme (ACE, kininase II) is a single-chain protein containing two active site domains (named N-and C-domains according to position in the chain). ACE is bound to plasma membranes by its C-terminal hydrophobic transmembrane anchor. Deal fluid, rich in ACE activity, obtained from patients after surgical colectomy was used as the source. Column chromatography, including modified affinity chromatography on lisinopril-Sepharose, yielded homogeneous ACE after only a 45-fold purification. N-terminal sequencing of ileal ACE and partial sequencing of CNBr fragments revealed the presence of an intact N terminus but only a single N-domain active site, ending between residues 443 and 559. Thus, ileal-fluid ACE is a unique enzyme differing from the widely distributed two-domain somatic enzyme or the single C-domain testicular (germinal) ACE. The molecular mass of ileal ACE is 108 kDa and when deglycosylated, the molecular mass is 68 kDa, indicating extensive glycosylation (37% by weight). In agreement with the results reported with recombinant variants of ACE, the ileal enzyme is less C1-dependent than somatic ACE; release of the C-terminal dipeptide from a peptide substrate was optimal in only 10 mM Cl. In addition to hydrolyzing at the C-terminal end of peptides, ileal ACE efficiently cleaved the protected N-terminal tripeptide from the luteinizing hormone-releasing hormone and its congener 6-31 times faster, depending on the Cl concentration, than the C-domain in recombinant testicular ACE. Thus we have isolated an active human ACE consisting of a single N-domain. We suggest that there is a bridge section of about 100 amino acids between the active N-and C-domains of somatic ACE where it may be proteolytically cleaved to liberate the active N-domain. These findings have potential relevance and importance in the therapeutic application of ACE inhibitors.

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Section: Resultsmentioning

confidence: 99%

Naturally occurring active N-domain of human angiotensin I-converting enzyme.

Deddish

Wang

Michel

et al. 1994

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

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“…Therefore, it is possible that some of the measured HL formation could be due to other peptidase activities in the gut, a tissue which is specialized in digesting proteins and other nutrients. On the other hand, early studies on intestinal ACE showed that three peaks of ACE activity could be separated and purified by Sepharyl gel filtration from human intestinal homogenates [36] and all of them could be inhibited by captopril. Diet can influence ACE activity in the intestinal epithelium.…”

Section: Discussionmentioning

confidence: 99%

Local Intestinal ACE-Like Activity and Corticosterone Production in Hypertensive and Aging Rats

Salmenkari¹,

Holappa²,

Siltari³

et al. 2018

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The renin angiotensin system (RAS) participates in inflammatory processes, as either a pro-inflammatory or an anti-inflammatory mediator. Components of RAS, such as angiotensin-converting enzyme (ACE), have been detected locally in the gut epithelium. In addition, the anti-inflammatory steroid, corticosterone, is produced in the gut. Hypertension and aging evoke a low-grade inflammatory process in the vascular endothelium. It is not known whether they induce a similar low-grade inflammation in the intestine and if the low-grade inflammation would evoke an activation of ACE and an elevation of corticosterone production. These two variables were measured in ileum and colon of 9-and 26-week old spontaneously hypertensive rats (SHR) and their normotensive Wistar-Kyoto (WK) controls. ACE-activity, measured via the formation of histidyl-leucine from hippuryl-histidyl-leucine in the tissue homogenate samples, was similar in the ileum and colon of young animals although the ileum of the young normotensive animals displayed the lowest level. In the old animals, the ACE activity was higher in the ileum than in the colon, especially in normotensive rats. Corticosterone production was measured as corticosterone concentration in the supernatants of ileum or colon after a 90-min ex vivo incubation. Corticosterone production was higher in ileum than in colon in both SHR and WK. No clear evidence was seen for age-dependency or for an effect of hypertension in the measured variables in the intestine. Thus, the putative low-grade inflammation in the intestine in aging or hypertension is not a strong enough stimulus to elevate corticosterone production or activate ACE.

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Local Intestinal ACE-Like Activity and Corticosterone Production in Hypertensive and Aging Rats

Salmenkari¹,

Holappa²,

Siltari³

et al. 2018

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Purification of Angiotensin-Converting Enzyme from Human Intestine

Cited by 3 publications

References 15 publications

Naturally occurring active N-domain of human angiotensin I-converting enzyme.

Naturally occurring active N-domain of human angiotensin I-converting enzyme.

Local Intestinal ACE-Like Activity and Corticosterone Production in Hypertensive and Aging Rats

Local Intestinal ACE-Like Activity and Corticosterone Production in Hypertensive and Aging Rats

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