Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.24.11 (CD10)
 9,792 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Degradation of Boc-CCK27-33 [Boc-Tyr(SO3H)-Met-Gly-Trp-Met-Asp-PheNH2) a fully potent analog of CCK8 was studied on synaptic plasma membranes from pig brain cortex. Characterization of the metabolites was performed by HPLC. This allowed to show the hydrolysis of the Asp-Phe bond by the neutral endopeptidase "enkephalinase", and the cleavages at the Met-Gly and Trp-Met bonds by PCMB sensitive enzymes. Similar results were observed using Boc(diNle28,31)-CCK27-33 as substrate. To investigate the biological relevance of these enzymes in the CCK8 metabolism, the degradation studies were performed on rat brain slices, with [3H]Boc(diNle28,31)CCK27-33 as substrate. Using these more physiological preparations i.e. striatal or cortex slices, the tritiated probe was cleaved at the Nle-Gly and Gly-Trp bonds. These degradation pathways were almost completely inhibited by PCMB, but in the striatum this inhibition process induced the appearance of a small peak corresponding to the action of enkephalinase. Taken together these results seem to indicate that thiol proteases play a crucial role in the CCK8 metabolism but that enkephalinase is virtually not involved.
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PMID:Investigation on the metabolism of CCK8 analogues by rat brain slices. 345 92

The complete amino acid sequence of the plastocyanin from the green alga Enteromorpha prolifera has been determined by Edman degradation of the intact molecule and fragments produced by enzymatic cleavage of the polypeptide chain with chymotrypsin, Staphylococcus aureus protease, proline-specific endopeptidase, Lys-C endopeptidase and trypsin. The molecule consists of 98 amino acid residues with a calculated relative molecular mass of 10103. The amino acid sequence of E. prolifera plastocyanin shows a high degree of homology with those plastocyanins from other algae and higher plants. In particular, the four residues which are copper ligands in other plastocyanins and in the bacterial electron transport protein azurin (two histidines, one cysteine and one methionine) are conserved. Five out of the six acidic amino acid side-chains which create an 'acidic patch' on the surface of plastocyanin from Populus nigra var. italica [Colman, P. M. et al. (1978) Nature (Lond.) 272, 319-324] are conserved in the amino acid sequence of E. prolifera plastocyanin.
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PMID:Complete amino acid sequence of plastocyanin from a green alga, Enteromorpha prolifera. 352 27

The effect of the arginine-specific reagents phenylglyoxal and butanedione on the activity of neutral endopeptidase 24.11 ("enkephalinase") was determined. Inactivation of the enzyme by butanedione is completely protected by methionine-enkephalin, but only partially protected by methionine-enkephalinamide. In contrast, phenylglyoxal inactivation of the enzyme exhibits saturation kinetics with a Kd of 20 mM. The enzyme is only partially protected against phenylglyoxal inactivation by both methionine-enkephalin and its amide, indicating that phenylglyoxal reacts at two sites. Reaction of the enzyme with phenylglyoxal in the presence of saturating methionine-enkephalin involves the direct reaction of the reagent with the enzyme-substrate complex. Enzyme treated with butanedione or with phenylglyoxal (at site 1) exhibits a 3-5 decrease in substrate binding with little change in kcat. In contrast, reaction with phenylglyoxal in the presence of saturating methionine-enkephalin shows little change in substrate binding but a 4-fold decrease in kcat. Enzyme inactivation involves the incorporation of approximately 1 mol of phenylglyoxal/enzyme subunit in the absence of methionine-enkephalin and approximately 2.5 mol of phenylglyoxal/enzyme subunit in the presence of saturating methionine-enkephalin. These results suggest that an arginine residue on the enzyme is involved in substrate binding.
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PMID:Reaction of neutral endopeptidase 24.11 (enkephalinase) with arginine reagents. 352 76

Tyr-Gly-Gly (YGG) was recently shown to be an extraneuronal metabolite of opioid peptides derived from proenkephalin A, formed in brain by the action of "enkephalinase" (membrane metalloendopeptidase, EC 3.4.24.11) and degraded by aminopeptidases. The dynamic state of YGG in mouse striatum was studied by evaluating the changes in its level elicited by inhibitors of these peptidases. Inhibition of YGG synthesis by Thiorphan or acetorphan reduced YGG levels with a t1/2 (mean +/- SEM) of 12 +/- 2 min, indicating an apparent turnover rate (mean +/- SEM) of 18 +/- 2 pmol/mg of protein per hr. An apparent turnover rate of 18 +/- 2 pmol/mg of protein per hr was derived from the rate of YGG accumulation elicited by the aminopeptidase inhibitor bestatin. In addition, accumulation of Tyr-Gly-Gly-Phe-Met (YGGFM) in an extrasynaptosomal fraction after blockade of its degradation by Thiorphan and bestatin occurred at a rate of 18 +/- 3 pmol/mg of protein per hr, which is likely to reflect the rate of enkephalin release in vivo. Hence, the three series of data suggest that striatal enkephalins rapidly turn over--e.g., with a t1/2 in the 1-hr range. Pentobarbital anesthesia reduced by about 60% the rate of YGG accumulation elicited by bestatin and the extrasynaptosomal YGGFM accumulation elicited by Thiorphan and bestatin. This suggests that the activity of striatal enkephalin neurons is depressed during anesthesia. Pentobarbital (and chloral hydrate) did not affect the steady-state level of YGGFM but rapidly reduced that of YGG. Hence, the steady-state levels of YGG seem a reliable index of changes in enkephalin release, and measuring levels of characteristic fragments might therefore provide a general means of evaluating neuropeptide release in vivo.
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PMID:Steady-state level and turnover rate of the tripeptide Tyr-Gly-Gly as indexes of striatal enkephalin release in vivo and their reduction during pentobarbital anesthesia. 352 54

Purified rat brain cathepsin B (EC 3.4.22.1) converted prodynorphins or proenkephalins to shorter active forms by the preferential removal of C-terminal dipeptides. The substrate affinities for Met-enkephalin-Arg-Phe or -Arg-Gly-Leu were Km 46 and 117 microM, and kcat/Km ratios were 67 and 115 microM-1, min-1, respectively. Met-Enkephalin was inactivated by the same mechanism (Km-450 microM; kcat/Km = 0.12 microM-1 min-1). The comparison of cathepsin B hydrolysis for pro-opioids, a synthetic hexapeptide and its fragments, C-blocked peptides (pro-opioid amides, Met-enkephalin amide, substance P), and bovine myelin basic protein, provided information on the influence of the C-terminal residues on dipeptide release, the rates as correlated to peptide length, and the optimal arrangement of residues favoring scission at the P1-P'1 sites. The brain enzyme was stereospecific and did not act on peptides with C-terminal D-amino acid substituents. Arg hindered and Pro blocked the release of C-terminal dipeptides when in the P'2 positions. The suppression of dipeptide release by agents inhibiting endopeptidase actions such as E-64 and leupeptin, and the endogenous brain factor (cerebrocystatin) point to similar catalytic mechanisms for the exopeptidase action.
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PMID:Preferential action of rat brain cathepsin B as a peptidyl dipeptidase converting pro-opioid oligopeptides. 353 Jan 35

Using ion-exchange chromatography on QAE-Sephadex A-50, affinity chromatography on DNP-hexamethylenediamine-Sepharose and gramicidin S-Sepharose and gel filtration, a metalloproteinase was isolated from the cultural fluid of L. pneumophila (strain Philadelphia-1) grown for 20 hours. The enzyme was purified 1606-fold with a 31% yield. The enzyme has a Mr of 38,000, pI approximately 4.0 and optimum of proteolytic activity at pH 6.0-7.0, 55 degrees C. The proteinase is the most stable within the pH range of 6.0-9.0. The enzyme contains one atom of zinc per molecule. The amino acid composition of metalloproteinase is close to that of thermolysin and is characterized by a high methionine content--17 residues out of 348. In the B-chain of oxidized bovine insulin the enzyme hydrolyzes the bonds precedent to the amino groups of leucine, phenylalanine and tyrosine. The enzyme is inhibited by chelating agents--Na2-EDTA and o-phenanthroline as well as by diethylpyrocarbonate. The serine and thiol proteinase inhibitors do not influence the enzyme activity. Under the given conditions of cultivation metalloproteinase is the major endopeptidase produced by L. pneumophila. Thus, the proteolytic system of Legionelles is characterized by the combination of metalloproteinase and the earlier described phenylalanine aminopeptidase.
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PMID:[Extracellular metalloproteinase from Legionella pneumophila]. 366 68

In some patients with genetic forms of extreme insulin resistance, there is a marked decrease in the number of insulin receptors on the cell surface. We studied an insulin-resistant patient (RM-1) with the Rabson-Mendenhall syndrome. As judged by insulin-binding studies, Epstein-Barr virus-transformed lymphocytes from patient RM-1 exhibit a 90% decrease in the number of insulin receptors. Similarly, with either lactoperoxidase-catalyzed radioiodination of cell surface receptors or biosynthetic labeling of receptors with [3H]glucosamine, we demonstrated an 80-90% decrease in the number of insulin receptors in cells from patient RM-1. Previous studies have shown that the marked decrease in insulin receptors of the Rabson-Mendenhall patient is not due to accelerated receptor degradation. Therefore, we investigated the possibility that a slow rate of receptor biosynthesis might account for the 90% reduction of insulin receptors in cells from this patient. Insulin-receptor biosynthesis proceeds through a glycoprotein precursor with an apparent Mr of 190,000. It undergoes endopeptidase cleavage and further posttranslational processing to yield the mature 135,000- and 95,000-Mr glycoprotein subunits. We studied the biosynthesis of the 190,000-Mr precursor and mature receptor subunits by a pulse-chase labeling technique with [2-3H]mannose. The time course of insulin-receptor biosynthesis appeared normal in cells from patient RM-1, despite a 10-fold reduction in the number of receptors on the cell surface. Parallel pulse-chase experiments with either [2-3H]mannose or [35S]methionine yielded the same results regardless of which label was employed. Thus, the receptor precursor in the Rabson-Mendenhall patient seems to be synthesized at a normal rat.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Insulin-receptor biosynthesis in cultured lymphocytes from an insulin-resistant patient (Rabson-Mendenhall syndrome). Evidence for defect before insertion of receptor into plasma membrane. 372 Oct 65

The secretions (HPS) contained an arylamidase-like enzyme discovered by chromatography on Sephadex G-100 Superfine columns using N-L-alanyl-2-naphthylamine (2NA) as substrate. The enzyme was fractionated in the void volume, suggesting that its molecular weight was 150,000 or higher. It hydrolysed, with decreasing rates, the 2NA of L-alanine, L-leucine, L-methionine and L-phenylalanine, the pH optimum for the best substrate (ala-2NA) being 8.0, alpha-Benzoyl-DL-arginine-2NA was not hydrolysed. p-Chloromercuribenzoate, EDTA, Ca2+ and Zn2+ were inhibitory, whereas chemical modification with typical tyrosyl group reagents did not significantly inactivate the enzyme. Treatment of HPS with Triton X-100 revealed two further arylamidase-like enzymes with lower mol. wt (90,000 and 40,000, respectively). Inhibition characteristics and Cl- effects suggest that one of these enzymes resembles aminopeptidase B (EC 3.4.11.6). HPS also contains endopeptidase activity over a wide pH range (6-9). The number of enzymes in HPS is thus small and most of the peptidolytic activity of HPS in vitro is due to one major enzyme with arylamidase activity.
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PMID:Partial purification and characterization of arylamidases from human palatine secretions. 386

Rat kidney membranes were solubilized by Triton X-100 and the CCK-8 degrading peptidases were resolved by chromatography on DEAE-cellulose. Four proteases were detected: two phosphoramidon-sensitive endopeptidases (EC 3.4.24.11), a bestatin-sensitive aminopeptidase and an unidentified enzyme. The pattern of cleavage of CCK-8 and shorter C-terminal fragments by endopeptidase 24.11 was investigated and indicated that the Gly29-Trp30, Trp30-Met31 and Asp32-Phe33 were scissile bonds. However, the cleavage pattern differed markedly from one CCK peptide to another: in the penta- and hexapeptide of CCK the bonds hydrolyzed were either Asp-Phe and Trp-Met or, Asp-Phe and Gly-Trp, respectively. The presence of the sulfate group on the tyrosine residue of CCK-8 influence markedly the nature of the major cleavage fragments produced by the endopeptidase. The major bonds cleaved were Asp-Phe, Trp-Met and Gly-Trp for unsulfated CCK-8, whilst for the sulfated octapeptide, the Trp-Met bond became a minor cleavage site.
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PMID:Hydrolysis of the C-terminal octapeptide of cholecystokinin by rat kidney membranes: characterization of the cleavage by solubilized endopeptidase-24.11. 389 58

The nerve growth factor dimer (beta NGF) can undergo two proteolytic modifications, one near the amino terminus where a unique histidine/methionine bond is cleaved and the other at the carboxy terminus releasing the terminal arginine residue. The modification near the amino terminus, releasing the first eight amino acids as an octapeptide, occurs as the result of the action of a specific endopeptidase present in the submaxillary gland. This same enzyme was found to be present in high concentration in epinephrine-stimulated saliva (20 mM) and was isolated from this source. The subunit interactions in 7S NGF were considered by assessing the protection afforded by each subunit, individually and in combination, against these two proteolytic modifications. Similar experiments were attempted with high molecular weight epidermal growth factor (HMW-EGF), an analogous growth factor complex that can experience a single modification, the release of arginine by carboxypeptidase B from the carboxy terminus of its biological subunit, low molecular weight EGF (LMW-EGF). The amino terminal octapeptide of beta NGF is protected by both the alpha and gamma subunits of 7S NGF and its loss has no effect on 7S NGF complex stabilization by zinc. The carboxy terminal arginine is protected only by the gamma subunit. A scheme depicting the subunit interactions in 7S NGF is presented.
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PMID:Subunit interactions of the nerve and epidermal growth factor complexes: protection of the biological subunit from proteolytic modification. 391 81


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