Expression of angiotensin I-converting enzyme in the human gastric HGT-1 cell line

Nonotte, Isabelle; Laliberté, Marie-France; Remy-Heintz, Nicole; Laliberté, François; Chevillard, Claude

doi:10.1016/0167-0115(95)00090-x

Cited by 6 publications

(3 citation statements)

References 19 publications

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“…However, it was expected, because we and others demonstrated previously that catalytically active ACE localized on the cell surface represents only part of the total ACE produced by cells [6], [46]–[47]. The remaining ACE protein in cells represents immature ACE, perhaps still containing the signal peptide [40], is catalytically inactive [6], [47], perhaps due to improper folding. Western blotting, which was performed with mAbs recognizing denatured epitopes (Fig.…”

Section: Resultsmentioning

confidence: 96%

Angiotensin I-Converting Enzyme Gln1069Arg Mutation Impairs Trafficking to the Cell Surface Resulting in Selective Denaturation of the C-Domain

et al. 2010

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BackgroundAngiotensin-converting enzyme (ACE; Kininase II; CD143) hydrolyzes small peptides such as angiotensin I, bradykinin, substance P, LH-RH and several others and thus plays a key role in blood pressure regulation and vascular remodeling. Complete absence of ACE in humans leads to renal tubular dysgenesis (RTD), a severe disorder of renal tubule development characterized by persistent fetal anuria and perinatal death.Methodology/Principal FindingsPatient with RTD in Lisbon, Portugal, maintained by peritoneal dialysis since birth, was found to have a homozygous substitution of Arg for Glu at position 1069 in the C-terminal domain of ACE (Q1069R) resulting in absence of plasma ACE activity; both parents and a brother who are heterozygous carriers of this mutation had exactly half-normal plasma ACE activity compared to healthy individuals. We hypothesized that the Q1069R substitution impaired ACE trafficking to the cell surface and led to accumulation of catalytically inactive ACE in the cell cytoplasm. CHO cells expressing wild-type (WT) vs. Q1069R-ACE demonstrated the mutant accumulates intracellularly and also that it is significantly degraded by intracellular proteases. Q1069R-ACE retained catalytic and immunological characteristics of WT-ACE N domain whereas it had 10–20% of the nativity of the WT-ACE C domain. A combination of chemical (sodium butyrate) or pharmacological (ACE inhibitor) chaperones with proteasome inhibitors (MG 132 or bortezomib) significantly restored trafficking of Q1069R-ACE to the cell surface and increased ACE activity in the cell culture media 4-fold.Conclusions/SignificanceHomozygous Q1069R substitution results in an ACE trafficking and processing defect which can be rescued, at least in cell culture, by a combination of chaperones and proteasome inhibitors. Further studies are required to determine whether similar treatment of individuals with this ACE mutation would provide therapeutic benefits such as concentration of primary urine.

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Section: Resultsmentioning

confidence: 96%

Angiotensin I-Converting Enzyme Gln1069Arg Mutation Impairs Trafficking to the Cell Surface Resulting in Selective Denaturation of the C-Domain

et al. 2010

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“…ACE, aminopeptidase N, and aminopeptidase W were detected both on the surface and in an intracellular pool in Caco-2 cells (15). Recently, the intracytoplasmic localization of ACE in the human gastric HGT-1 cell line has been found; however, most of the ACE within these ceils was immuuologically active, but enzymatically inactive (24). Taken together, these studies demonstrate the existence of an intracellular pool of ACE in some cell types, but a similar result has not been previously reported for HUVEC.…”

Section: Discussionmentioning

confidence: 97%

Modulation of angiotensin-converting enzyme in cultured human vascular endothelial cells

Balyasnikova

Danilov

Muzykantov

et al. 1998

In Vitro Cell.Dev.Biol.-Animal

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Previous work has suggested that not all immunoreactive angiotensin-converting enzyme (ACE) in tissues or cells is in a biologically active state. We have explored this possibility in cultured human umbilical vein endothelial cells (HUVEC), one of the most widely studied in vitro endothelial cell systems. Our approach included characterization of the effect of increasing passage number on ACE activity and expression of immunoreactive ACE at the single cell level, the subcellular compartmentalization of active ACE, and the effect of phorbol ester (PMA) treatment. We found that both ACE activity and expression of ACE antigen were downregulated by cultivation (30% of ACE-positive cells at seventh passage vs. 90% in primary culture). ACE downregulation is specific (number of CD31-positive cells did not change with cultivation) and correlated with downregulation of factor VIII-antigen. The percentage of ACE-positive cells in permeabilized HUVEC at third passage was almost twice that in nonpermeabilized HUVEC (90% vs. 50%), indicating that HUVEC contain intracellular immunoreactive ACE. ACE activity, however, was similar when measured in intact cells and in cell lysates. Moreover, diazonium salt of sulfanilic acid (DASA), a membrane-impermeable ACE inhibitor, inhibited ACE activity in intact cells and in cell lysates at the same extent, thus implying that intracellular ACE is inactive. PMA (100 nM) treatment increased the percentage of ACE-positive cells at third passage from 57 to 96%. ACE activity was increased 3-fold in cell and 1.5-fold in the culture medium of PMA-treated cells. Analysis of ACE activity in intact monolayers and cell lysates of control and PMA-treated cells revealed that all enzymatically active ACE in PMA-treated cells is localized on the plasma membrane and acts as an ectoenzyme. We conclude that expression of ACE by HUVEC is downregulated by repeated passage in culture but can be restored by PMA treatment. In addition, ACE expression is heterogeneous between neighboring cells, and total immunoreactive ACE protein associated with HUVEC includes an inactive pool of the enzyme.

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“…HGT-1 cells have been shown to express histamine H 2 receptors, which have been demonstrated to mediate adenylyl cyclase activation and cAMP production [ 30 ]. Furthermore, this cell line has been shown to express acetylcholine receptors, the H + /K + ATPase proton pump, an omeprazol-binding site possessing a K + channel [ 31 ], angiotensin 1-converting enzyme [ 32 ], and transporters required for gastric acid secretion [ 33 ]. HGT-1 cells have consequently been established as a valuable cell model for assessing the effects of different food constituents, found, e.g., in coffee beverages, on proton secretion by measuring the intracellular pH value [ 34 , 35 ].…”

Section: Introductionmentioning

confidence: 99%

Gastric Serotonin Biosynthesis and Its Functional Role in L-Arginine-Induced Gastric Proton Secretion

Holik

Schweiger

Stoeger

et al. 2021

IJMS

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Among mammals, serotonin is predominantly found in the gastrointestinal tract, where it has been shown to participate in pathway-regulating satiation. For the stomach, vascular serotonin release induced by gastric distension is thought to chiefly contribute to satiation after food intake. However, little information is available on the capability of gastric cells to synthesize, release and respond to serotonin by functional changes of mechanisms regulating gastric acid secretion. We investigated whether human gastric cells are capable of serotonin synthesis and release. First, HGT-1 cells, derived from a human adenocarcinoma of the stomach, and human stomach specimens were immunostained positive for serotonin. In HGT-1 cells, incubation with the tryptophan hydroxylase inhibitor p-chlorophenylalanine reduced the mean serotonin-induced fluorescence signal intensity by 27%. Serotonin release of 147 ± 18%, compared to control HGT-1 cells (set to 100%) was demonstrated after treatment with 30 mM of the satiating amino acid L-Arg. Granisetron, a 5-HT3 receptor antagonist, reduced this L-Arg-induced serotonin release, as well as L-Arg-induced proton secretion. Similarly to the in vitro experiment, human antrum samples released serotonin upon incubation with 10 mM L-Arg. Overall, our data suggest that human parietal cells in culture, as well as from the gastric antrum, synthesize serotonin and release it after treatment with L-Arg via an HTR3-related mechanism. Moreover, we suggest not only gastric distension but also gastric acid secretion to result in peripheral serotonin release.

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Expression of angiotensin I-converting enzyme in the human gastric HGT-1 cell line

Cited by 6 publications

References 19 publications

Angiotensin I-Converting Enzyme Gln1069Arg Mutation Impairs Trafficking to the Cell Surface Resulting in Selective Denaturation of the C-Domain

Angiotensin I-Converting Enzyme Gln1069Arg Mutation Impairs Trafficking to the Cell Surface Resulting in Selective Denaturation of the C-Domain

Modulation of angiotensin-converting enzyme in cultured human vascular endothelial cells

Gastric Serotonin Biosynthesis and Its Functional Role in L-Arginine-Induced Gastric Proton Secretion

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