Distinction between endo-oligopeptidase A (EC 3.4.22.19) and soluble metalloendopeptidase (EC 3.4.24.15)

Camargo, Antônio Carlos Martins de

doi:10.1042/bj2770294b

Cited by 8 publications

(10 citation statements)

References 18 publications

(21 reference statements)

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“…The acronym 'thimct oligopeptidase' (thiol-and metal-dependent oligopeptidase) has been introduced for these enzymes [26,291. However, the identity of endo-oligopeptidase A (or Pz-peptidase) with endopeptidase 24.15 has been opposed [30,31]. In our opinion at least two (endopeptidases 24.15 and 24.16) and probably more (endo-oligopeptidase A/Pz-peptidase) proteases with similiar molecular and catalytic properties exist which are all thiol-and metal-dependent enzymes.…”

Section: Inhibitors and Activators Discussionmentioning

confidence: 97%

Purification of the main somatostatin‐degrading proteases from rat and pig brains, their action on other neuropeptides, and their identification as endopeptidases 24.15 and 24.16

Dahms

Mentlein

1992

European Journal of Biochemistry

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The main somatostatin-degrading proteases were purified from rat and pig brain homogenates and characterized as thiol-and metal-dependent endoproteases. Two types of proteases with apparent native and subunit molecular masses of 70 kDa and 68 kDa could be differentiated in both species. Beside somatostatin, both hydrolyzed several other neuropeptides with chain lengths between 8 and 30 amino acid residues. Cleavage sites were generally similar or identical, but some clear exceptions were observed for enzymes from both species which could be used to differentiate between the two proteases. The 68-kDa protease cleaved somatostatin at three bonds (Asn5-Phe6, Phe6-Phe7 and ThrlO-Phell) and neurotensin only at the Arg8-Arg9 bond, whereas the 70-kDa protease digested somatostatin at only two bonds (Phe6-Phe7 and ThrlO-Phel 1) and neurotensin as well as acetylneurotensin-(8 -13) additionally (pig protease) or almost exclusively (rat protease) at the ProlOTyrll bond. Relative rates for the digestions of various peptides were, however, more dependent on the species than on the type of protease. Cleavage sites for angiotensin 11, bradykinin, dynorphin, gonadoliberin and substance P were, apart from different rates, identical for both proteases. In both species the 68-kDa protease was found to be mainly, but not exclusively, soluble and not membraneassociated, whereas the inverse was detected for the 70-kDa protease. Based on distinct molecular and catalytic properties, the 68-kDa protease is supposed to be congruent with the endopeptidase 24.15 (EC 3.4.24.15), the 70-kDa protease with endopeptidase 24.16 (EC 3.4.24.16, neurotensindegrading endopeptidase). This investigation demonstrates that both proteases hydrolyze various neuropeptides with similar cleavage sites. but with species-dependent activity. Species-independent distinctions are the exclusive action of endopeptidase 24.16 on acetylneurotensin-(8 -13) and liberation of free Phe from somatostatin only by endopeptidase 24.15.The cyclic tetradecapeptide somatostatin, originally purified as an inhibitory factor for the release of somatotropin from pituitary cells [l], is a widespread neurotransmitter or neuromodulator in the brain [2]. Somatostatin Enzymes. Soluble mctallo-endopeptidase (EC 3.4.24.1 5); neurotensin-degrading neutral metallopeptidase (EC 3.4.24.16); endo-oligopeptidase (EC 3.4.22.19); tissue-endopeptidase degrading collagenase-synthetic substrate, Pz-peptidase (EC 3.4.99.31); enkephalinase, membrane metallo-endopeptidase (EC 3.4.24.1 1); prolyl endopeptidase (thiol-dependent) (EC 3.4.22.18); cytosol aminopeptidase, leucyl aminopeptidase (EC 3.4.1 1.1); aminopeptidase M or N, microsomal (alanine) aminopeptidase (EC 3.4.1 1.2). vated to avoid a constant stimulus. However, as for other neuropeptides, the factors responsible for inactivation in the brain are insufficiently known [8 -lo].Recently, we have shown that somatostatin is inactivated by proteolytic degradation after exposure to cultivated neuronal and glial cells from rat brain [ll]. Hydrolysis...

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Section: Inhibitors and Activators Discussionmentioning

confidence: 97%

Purification of the main somatostatin‐degrading proteases from rat and pig brains, their action on other neuropeptides, and their identification as endopeptidases 24.15 and 24.16

Dahms

Mentlein

1992

European Journal of Biochemistry

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“…A similar enzyme was also described in rat brain and classified as a metalloendopeptidase (MEP 24.15,EC 3.4.24.15) [Orlowski et al, 19831. The identity of these two enzymes is still controversial and awaiting confirmation [Camargo, 1991;Barret, 1991;Gomes et al, 19931, once both enzymes are able to hydrolyse and inactivate bioactive peptides such as bradykmin, neurotensin, and luteinizing hormone releasing hormone (LHRH) [Oliveira et al, 1976;Carvalho and Camargo, 1981;Camargo et al, 1982Camargo et al, , 1984Chu and Orlowski, 1985;Orlowski et al, 19891. In addition, they convert by single cleavage small enkephalin-containing peptides (8-13 amino acidis residues) into [Leu5]-enkephalin or LMet51-enkephalin [Camargo et al, 1985[Camargo et al, , 1987Chu and Orlowski, 19851. Using PC12 cells as a promising in vitro cellular model to study bioactive peptide metabolism, we described here the preliminary characterization of an EOPA-like activity in PC12 cells and we also showed evidence that cyclic AMP analogs but not FGF modulate this activity.…”

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confidence: 98%

Characterization of an endooligopeptidase A‐like protein in PC12 cells: Activity modulation by cAMP but not by basic fibroblast growth factor

Ferro

Tambourgy

Abreu

et al. 1995

J of Cellular Biochemistry

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Endooligopeptidase A is a putative neuropeptide-metabolizing enzyme. It converts small enkephalin-containing peptides into the corresponding enkephalins and inactivates biopeptides such as bradykinin and neurotensin in vitro. We investigated the presence of endooligopeptidase A in PC12 cells. This cell line was derived from a rat pheochromocytoma tumor and resembles fetal chromaffin cell. Depending on the supplements added to the cell culture, this cell line can be differentiated into mature chromaffin cell or sympathetic neuron-like cell. Endooligopeptidase A activity was measured in soluble cellular extracts using a specific fluorogenic substrate QF-ERP7. The PC12 endooligopeptidase A-like activity shared similar but not identical biochemical properties with rabbit brain endooligopeptidase A. Similarly to rabbit brain endooligopeptidase A, the PC12 endooligopeptidase A-like activity was enhanced by DTT, totally inhibited by DTNB and 1-10 Phenanthroline, partially inhibited by cFP-AAF-pAb, and not affected by PMSF. Furthermore, the PC12 endooligopeptidase A-like activity displayed identical elution profile as rabbit brain endooligopeptidase A in gel filtration and anion-exchange chromatography. In addition, an antiserum raised against rabbit brain endooligopeptidase A cross-reacted with a 71 kDa component from PC12 cell extracts in Western blotting and was also able to partially neutralize the PC12 endooligopeptidase A-like activity. Treatment of PC12 cells with basic fibroblast growth factor (bFGF), a neurotrophic factor for this cell line, did not modify the specific activity of this enzyme. However, cAMP analogs decreased the specific activity of the enzyme. These results indicate the presence of an endooligopeptidase A-like activity in PC12 cells which is modulated by cAMP but not by bFGF.

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“…However, the presence of a cysteine residue close to the catalytic centre of endopeptidase-24.15 might explain the activation of the enzyme by 2-mercaptoethanol and its inhibition by pmercuribenzoate and TV-ethylmaleimide. Although it has been claimed that endo-oligopeptidase A and endopeptidase-24.15 may be identical (Barrett & Brown, 1990), an antibody to endo-oligopeptidase A does not precipitate endopeptidase-24.15 (Toffoletto, Metters, Oliveira et al 1988) and the two activities are separable by ion exchange chromatography (Camargo, 1991 ). The relationship between these two activities remains unresolved, therefore, and must await the cloning and sequencing of endo-oligopep¬ tidase A.…”

Section: Characterization Of Human Endo-oligopeptidases a Andmentioning

confidence: 98%

“…Endo-oligopeptidase A resembles in specificity another endopeptidase, now designated endopeptidase-24.15 (EC 3.4.24.15), which is highly active in rat brain and pituitary (Orlowski, Michaud & Chu, 1983) and which has recently been cloned and sequenced (Pierotti, Dong, Glucksman et al 1990). The conclusion that endopeptidase-24.15 is identical to endo-oligopeptidase A and to another activity, 4-phenylazobenzyloxycarbonyl-peptidase Tisljar, Camargo, Costa & ; Barrett & Brown, 1990) was later contested by Camargo (1991), who found that biochemical dis¬ tinctions can be made between endo-oligopeptidase A and endopeptidase-24.15, including physical separa¬ tion. The cytosol of nervous tissue also contains another thiol-activated endopeptidase named endooligopeptidase (Oliveira, Martins & Camargo, 1976) further characterized as prolyl-endopeptidase (Greene, Spadaro, Martins et al 1982) which hydro¬ lyses bradykinin at the Pro7-Phe8 bond (Oliveira et al 1976) as well as the carboxyl group of proline in a number of biologically active peptides (Orlowski, Wilk, Pearce & Wilk, 1979;Greene et al 1982).…”

Section: Introductionmentioning

confidence: 96%

Distribution and properties of endo-oligopeptidases A and B in the human neuroendocrine system

Medeiros

Iazigi²,

Camargo³

et al. 1992

Journal of Endocrinology

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The thiol activated endo-oligopeptidases A and B were studied in the soluble fraction of human hypothalamus and various endocrine glands. For the identification, characterization and purification of the enzymes, Z-Gly-Pro-NH-Np and bradykinin were used as substrates. Endo-oligopeptidase B showed a molecular mass ranging from 55.5 to 65.5 kDa and isoelectric point from 4.7 to 4.95. Its activity in tissues was highest in the testis, with intermediate levels in the thyroid, neurohypophysis, adenohypophysis and hypothalamus and the lowest activity in the pineal gland. Endo-oligopeptidase A, 467-fold purified by immunoaffinity chromatography, exhibited a molecular mass of 65.5 kDa in the adenohypophysis but 58.5 kDa in other tissues. The isoelectric point ranged from 5.22 to 5.50. High endo-oligopeptidase A activity was observed in the adenohypophysis, testis and hypothalamus with lesser activity in the neurohypophysis and thyroid and the lowest in the pineal gland. Endo-oligopeptidase A cleaved the bonds Phe-Ser of bradykinin, Met-Arg of BAM-12P and Arg-Arg of neurotensin as described for rabbit brain and heart and bovine brain enzymes. This work shows that endo-oligopeptidase A also hydrolysed the bonds Tyr-Gly of LH-releasing hormone, Pro-Phe of angiotensin I and Tyr-Ile of angiotensin II.

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Distinction between endo-oligopeptidase A (EC 3.4.22.19) and soluble metalloendopeptidase (EC 3.4.24.15)

Cited by 8 publications

References 18 publications

Purification of the main somatostatin‐degrading proteases from rat and pig brains, their action on other neuropeptides, and their identification as endopeptidases 24.15 and 24.16

Purification of the main somatostatin‐degrading proteases from rat and pig brains, their action on other neuropeptides, and their identification as endopeptidases 24.15 and 24.16

Characterization of an endooligopeptidase A‐like protein in PC12 cells: Activity modulation by cAMP but not by basic fibroblast growth factor

Distribution and properties of endo-oligopeptidases A and B in the human neuroendocrine system

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