A highly sensitive fluorometric assay for “enkephalinase”, a neutral metalloendopeptidase that releases tyrosine-glycine-glycine from enkephalins

Florentin, D.; Sassi, Amor; Roques, Bernárd P.

doi:10.1016/0003-2697(84)90425-1

Cited by 114 publications

(52 citation statements)

References 23 publications

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“…The sample was centrifuged at 41C and plasma was stored at À801C. The NEP activity was measured fluorimetrically according to Florentin et al 19 Briefly, plasma (15 ml) was incubated at 371C (1-10 mmol/l) and decreasing concentrations of DAGPNG (50-40 mmol/l). Proteins were measured by the Bradford's method.…”

Section: Measurement Of Plasma Ace Activitymentioning

confidence: 99%

Neutral endopeptidase and angiotensin I converting enzyme insertion/deletion gene polymorphism in humans

Oliveri

et al. 2004

J Hum Hypertens

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Neutral endopeptidase (NEP) hydrolyses angiotensins (Ang) I and II and generates angiotensin-(1-7) ]. In humans, the insertion/deletion (I/D) angiotensin-I converting enzyme (ACE) gene polymorphism determined plasma ACE levels by 40%. In rats, a similar polymorphism determines ACE levels which are inversely associated to NEP activity. The objective of this study is to evaluate the relationship between ACE expression and plasma NEP activity in normotensive subjects and in hypertensive patients. In total, 58 consecutive patients with hypertension, evaluated in our Hypertension Clinic, were compared according to their ACE I/D genotypes with 54 control subjects in terms of both plasma ACE activity and NEP activities. Plasma ACE activity was elevated 51 and 70% in both DD ACE groups (normotensives and hypertensives) compared with their respective ID and II ACE groups (Po0.001). A significant effect of the ACE polymorphism and of the hypertensive status on ACE activity was observed (Po0.001). In normotensive DD ACE subjects, NEP activity was 0.30 7 0.02 U/ml, whereas in the normotensive II ACE and in the normotensive ID ACE subjects NEP activity was increased 65 and 48%, respectively (Po0.001). In the hypertensive DD ACE patients, NEP activity was 0.47 7 0.03 U/mg. An effect of the I/D ACE genotypes on NEP activity (Po0.04) and an interaction effect between the I/D ACE genotype and the hypertensive status were also observed (Po0.001). These results are consistent with a normal and inverse relationship between the ACE polymorphism and NEP activity in normotensive humans (as is also observed in rats). This normal relationship is not observed in hypertensive patients.

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Section: Measurement Of Plasma Ace Activitymentioning

confidence: 99%

Neutral endopeptidase and angiotensin I converting enzyme insertion/deletion gene polymorphism in humans

Oliveri

et al. 2004

J Hum Hypertens

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“…22 Na in Earle's salt solution, 25 mM Hepes-Tris (pH 7.4) at 37°C for 3 min with A7r5 cells 43) ; adenylate cyclase: ATP and forskolin in 25 mM Tris-HCl buffer (pH 7.5) at 30°C for 30 min with rat brain 44) ; guanylate cyclase: GTP and SNP in 40 mM triethanolamine (TEA)-HCl buffer (pH 7.4) at 30°C for 15 min with bovine lung 45) ; b-adrenoceptor-G protein coupling (agonist effect): rat heart cells in 25 mM Tris-HCl buffer (pH 7.5) at 30°C for 30 min 44) ; acetylcholinesterase: AMTCh in 0. ; neutral endopeptidase: DAGNPG in 50 mM Tris-HCl buffer (pH 7.4) at 37°C for 30 min with neutral endopeptidase 54) ; MMP-3: NFF-2 in 50 mM Tris-HCl buffer (pH 7.5) at 37°C for 90 min with MMP-3 55) ; MMP-7: MMP-2/MMP-7 substrate in 50 mM Tris-HCl buffer (pH 7.5) at 37°C for 45 min with MMP-3 56) ; MMP-9: NFF-2 in 50 mM Tris-HCl buffer (pH 7.5) at 37°C for 90 min with MMP-9 55) ; cell proliferation (serum-stimulated A7r5): A7r5 cells in DMEM at 37°C for 24 h …”

Section: Animals and Anesthesiamentioning

confidence: 99%

Effects of the New Ethacrynic Acid Derivative SA9000 on Intraocular Pressure in Cats and Monkeys

Shimazaki

Ichikawa

Rao

et al. 2004

Biological & Pharmaceutical Bulletin

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In glaucoma, the second most common cause of blindness, an elevated intraocular pressure (IOP) is one of the risk factors for damage to the optic nerve. Currently, application of ocular hypotensive drugs is the mainstream approach to glaucoma therapy, and extensive efforts have been made to develop anti-glaucoma drugs that lower IOP. These drugs are intended either to modulate aqueous humor outflow at sites in the trabecular meshwork or ciliary muscle, [1][2][3][4][5] or to inhibit the production of aqueous humor by the ciliary body. Aqueous humor outflow consists of conventional trabecular meshwork outflow and unconventional uveoscleral outflow. It has been proposed that TM plays the major role in the regulation of normal aqueous humor outflow to maintain normal range of IOP. [6][7][8][9] At present, many ocular hypotensive agents that modulate uveoscleral outflow (such as the prostaglandin derivatives) are in use as anti-glaucoma drugs. In contrast, there are no drugs that act directly on the trabecular meshwork to increase conventional outflow, even though this constitutes approximately 90% of the normal eye's total aqueous outflow. 10)Thus, an effective modulator of conventional outflow might be capable of exerting a powerful ocular hypotensive effect and be of great value as a next-generation ocular hypotensive drug.Ethacrynic acid (ECA) is well known as a sulfhydryl (SH) reactive diuretic and a NaϪ co-transport system inhibitor, 11,12) and it has been observed to increase aqueous humor outflow and lower IOP. ECA has been reported to increase aqueous humor outflow facility in enucleated animal and human eyes and in living monkey eyes, [13][14][15][16][17] and these effects have been correlated with changes in human trabecular meshwork cell shape and cytoskeleton in vitro.18) IOP lowering also has been observed in living human eyes with glaucoma. 19) ECA thus seems to have the potential to become an ocular hypotensive agent by its targeting of the trabecular meshwork, a site at which it seems to cause reversible changes in both cell shape and attachment.18) Nevertheless, because of its possible ocular side effects, there is a need for derivatives of ECA with even greater ocular safety 16,17) and corneal penetration. 20) To broad the therapeutic index, which is shown as the ratio of doses between the IOP reducing effects and potential side effects, may lead to find the clinically useful ocular hypotensive drugs. We, therefore, synthesized thirty-two compounds and evaluated ECA derivatives that might retain the strong cytoskeletal modulating activity of ECA while having a lower cytotoxicity potential, i.e. a broader therapeutic index. The structural modifications of ECA examined involved both the phenoxyacetic acid and the acryloyl moieties. Among these, we found a new ECA derivative, SA9000, which appeared to have a broader therapeutic index. In the present study, we examined the effects of SA9000 on IOP in cats and monkeys, and also evaluated changes in corneal endothelial and epithelial morphology as surrog...

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“…At increasing reaction times (0-120 min), 0. l-ml aliquots were quenched in ice-cold Pipes buffer containing 5 mM EDTA, centrifuged to remove the fine precipitate formed in the reaction, and processed as above. A doubling in the initial fluorescence intensity corresponds to the hydrolysis of 20% of the substrate (40). sequence of a CALLA cDNA clone is identical to that coding for human neutral endopeptidase (14).…”

Section: Introductionmentioning

confidence: 96%

“…Assays were performed essentially as described (40), using the synthetic substrate dansyl-D-AlaGly-Phe(pNO2)-Gly (DAGNPG; Sigma Chemical Co., St Louis, MO).…”

Section: Introductionmentioning

confidence: 99%

Common acute lymphoblastic leukemia antigen expressed on leukemia and melanoma cell lines has neutral endopeptidase activity.

Jongeneel

Quackenbush²,

Ronco³

et al. 1989

J. Clin. Invest.

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We have previously reported that the amino acid sequence of the common acute lymphoblastic leukemia antigen (CALLA, CD10) translated from a normal human kidney cDNA clone is identical to that of neutral endopeptidase (NEP, EC 3.4.24.11). In this study, we show that by flow cytometry, a monoclonal antibody (135A3) produced against rabbit NEP reacted selectively with leukemia and melanoma cell lines expressing CALLA on their surface. A glycoprotein of apparent

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A highly sensitive fluorometric assay for “enkephalinase”, a neutral metalloendopeptidase that releases tyrosine-glycine-glycine from enkephalins

Cited by 114 publications

References 23 publications

Neutral endopeptidase and angiotensin I converting enzyme insertion/deletion gene polymorphism in humans

Neutral endopeptidase and angiotensin I converting enzyme insertion/deletion gene polymorphism in humans

Effects of the New Ethacrynic Acid Derivative SA9000 on Intraocular Pressure in Cats and Monkeys

Common acute lymphoblastic leukemia antigen expressed on leukemia and melanoma cell lines has neutral endopeptidase activity.

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