Prolyl endopeptidase from bovine testis: Purification, characterization and comparison with the enzymes from other tissues.

Yoshimoto, Tadashi; Oyama, Hiroshi; Koriyama, Nobuhiro; Tsuru, Daisuke

doi:10.1248/cpb.36.1456

Cited by 11 publications

(9 citation statements)

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“…The molecular weight of the native enzyme was 74,000 as determined by gel filtration on Sephadex G-150, indicating that the enzyme is a monomer with a molecular weight of around 75,000. This value corresponds to the 68,000 to 77,000 previously determined for proline-specific enzymes from plants (21,37,38), mammalian tissues (10,20,23,36), and Flavobactenum meningosepticum (32,40). The isoelectric point was 6.2, as compared with 9.6 for the Flavobactenium enzyme (32).…”

Section: Resultssupporting

confidence: 68%

“…PPSE was inhibited by diisopropylfluorophosphate, like other proline-specific endopeptidases (10,20,21,23,30,(34)(35)(36)(37)40), and it may therefore be classified as a serine protease. The activity of PPSE appears not to be dependent on thiol groups, since it was not sensitive to mercuric ions.…”

Section: Resultsmentioning

confidence: 99%

“…These enzymes should be distinguished from the special-purpose endopeptidases (involved in, e.g., hormone processing and blood clotting) that recognize several amino acid residues in the substrate. Enzymes with specificity for cleavage at the carboxyl side of Asp and Glu (6,11,18), Glu (25,31), Lys (13), Arg (7), Val (1), Gly (5), and Pro (2,10,20,21,23,34,(36)(37)(38)40) have been described. These enzymes are of interest from the point of view of understanding the nature of their high specificity, but they have also been used for specific cleavage of fusion proteins and for synthesis of peptide bonds.…”

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confidence: 99%

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Proline-specific endopeptidases from microbial sources: isolation of an enzyme from a Xanthomonas sp

Rasmussen

Meldal

et al. 1992

J Bacteriol

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An extensive screening among microorganisms for the presence of post-proline-specific endopeptidase activity was performed. This activity was found among ordinary bacteria from soil samples but not among fungi and actinomycetes. This result is in contrast to the previous notion that this activity is confined to the genus Flavobacterium. A proline endopeptidase was isolated from a Xanthomonas sp. and characterized with respect to physicochemical and enzymatic properties. The enzyme is composed of a single peptide chain with a molecular weight of 75,000. The isoelectric point is 6.2. It is inhibited by diisopropylfluorophosphate and may therefore be classified as a serine endopeptidase. The activity profile is bell shaped with an optimum at pH 7.5.By using synthetic peptide substrates and intramolecular fluorescence quenching it was possible to study the influence of substrate structure on the rate of hydrolysis. The enzyme specifically hydrolyzed Pro-X peptide bonds. With Glu at position X, low rates of hydrolysis were observed; otherwise the enzyme exhibited little preference for particular amino acid residues at position X. A similar substrate preference was observed with respect to the amino acid residue preceding the prolyl residue in the substrate. The enzyme required a minimum of two amino acid residues toward the N terminus from the scissile bond, but further elongation of the peptide chain by up to six amino acid residues caused only a threefold increase in the rate of hydrolysis. Attempts to cleave at the prolyl residues in oxidized RNase failed, indicating that the enzyme does not hydrolyze long peptides, a peculiar property it shares with other proline-specific endopeptidases.Over the past 20 years a number of serine and thiol endopeptidases that cleave specifically on the carboxyl side of particular amino acid residues in peptides and proteins have been identified. These enzymes should be distinguished from the special-purpose endopeptidases (involved in, e.g., hormone processing and blood clotting) that recognize several amino acid residues in the substrate. Enzymes with specificity for cleavage at the carboxyl side of Asp and Glu (6,11,18), Glu (25, 31), Lys (13), Arg (7), Val (1), Gly (5), and Pro (2,10,20,21,23,34,(36)(37)(38)40) have been described. These enzymes are of interest from the point of view of understanding the nature of their high specificity, but they have also been used for specific cleavage of fusion proteins and for synthesis of peptide bonds.Due to the unusual structure of proline, most endopeptidases hydrolyze Pro-X bonds at extremely low rates; this has increased the interest in proline-specific enzymes. Such enzymes are commonly found in small quantities in plants (21, 37, 38) and in mammalian organs (2,10,20,23,29,34), where they appear to function in the degradation of numerous proline-containing peptide hormones (12,15,(26)(27)(28). In contrast, they have been reported to be rare in microorganisms; extensive screening only identified a single genus, Flavobacterium, with su...

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Section: Resultssupporting

confidence: 68%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Proline-specific endopeptidases from microbial sources: isolation of an enzyme from a Xanthomonas sp

Rasmussen

Meldal

et al. 1992

J Bacteriol

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“…S-17092-1 was kindly given by Prof. F. Checler. [36] Experiments with ZGP-pNA: POP activity was determined by the method described by Yoshimoto et al [37] Briefly, the reactions were performed in micro titer plates of 96 wells, which allowed the simultaneous monitoring of multiple reactions. For each reaction, activity buffer (131 mL,100 mm of Na/K phosphate buffer, pH 8.0) was pre-incubated for 15 min at 30 8C with hPOP (7 nm) and with the corresponding inhibitor solution (6 mL) or activity buffer (controls).…”

Section: Pop Activity Assaysmentioning

confidence: 99%

Identification by ¹⁹F NMR of Traditional Chinese Medicinal Plants Possessing Prolyl Oligopeptidase Inhibitory Activity

et al. 2006

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Prolyl oligopeptidase is a cytosolic serine peptidase that hydrolyzes proline-containing peptides at the carboxy termini of the proline residues. This peptidase has been associated with schizophrenia, bipolar affective disorder, and related neuropsychiatric disorders and might therefore have important clinical implications. Traditional Chinese medicinal (TCM) plants provide a rich source of unexplored compounds for strategies to find novel POP inhibitors, but the traditional methodologies used to identify POP inhibitors could have some limitations when working with natural products: interference with the colorimetric or fluorimetric detection methods commonly used to screen for POP inhibitors can result in the generation of false positives or false negatives. Since NMR screening is less prone to such interference, we decided to explore the use of 19F NMR to screen for POP inhibitors. We synthesized a new 19F-labeled POP substrate--Z-Gly-Pro-Phe-4(CF3)-NH2--and used it to search for new POP inhibitors in TCM plant extracts. We identified several plants with high POP-inhibitory activity and show here that the combination of 19F NMR and TCM plant extracts is a useful tool for identifying new POP inhibitors.

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“…First isolated in the human uterus as an oxytocin-degrading enzyme (Walter et al, 1971), PEP was subsequently purified from lamb kidney (Koida and Walter, 1976), and the PEP cDNAs have been cloned in human (Vanhoof et al, 1994), mouse (Ishino et al, 1998), and rat (Kimura and Takahashi, 2000). PEP is widely distributed in mammalian tissues, including muscle (Daly et al, 1985), liver (Yamakawa et al, 1986), testis (Yoshimoto et al, 1988), and brain (Kato et al, 1980;Kalwant and Porter, 1991). A crystal structure analysis of PEP has revealed that the catalytic site is covered by an unclosed seven-bladed ␤-propeller domain that is thought to exclude peptides longer than 30 amino acids and a fortiori proteins from proteolysis (Fü löp et al, 1998(Fü löp et al, , 2000.…”

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confidence: 99%

Localization of the mRNA encoding prolyl endopeptidase in the rat brain and pituitary

Bellemère

Vaudry

Mounien

et al. 2004

J of Comparative Neurology

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Prolyl endopeptidase (EC 3.4.21.26, PEP), a serine protease that hydrolyzes peptides at the carboxyl side of proline residues, is involved in the breakdown of several proline-containing neuropeptides and, thus, may contribute to the regulation of behavioral activities. In this study, the distribution of PEP mRNA was investigated in the central nervous system and pituitary of rat by means of quantitative reverse transcriptase-polymerase chain reaction analysis and in situ hybridization histochemistry. High densities of PEP transcripts were found in cerebellar Purkinje and granule cells, within most hypothalamic nuclei, in pyramidal neurons of the Ammon's horn, in granule cells of the dentate gyrus, and within the basolateral complex of the amygdala. Moderate levels of PEP mRNA were observed in layers 3-5 of the cerebral cortex, the anterior thalamic group, the septal region, the substantia nigra, the magnocellular neurons of the red nucleus, and the motor nuclei of the cranial nerves. Low concentrations of PEP mRNA were detected in the deep mesencephalic nuclei, the reticular formation, the pretectum, and the tectum. A high density of PEP mRNA was found in the intermediate and the anterior lobes of the pituitary, while the neural lobe was devoid of labeling. In several brain regions, the distribution pattern of PEP mRNA overlapped that of various neuropeptide receptors, suggesting that PEP is actually involved in the inactivation of regulatory neuropeptides.

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Prolyl endopeptidase from bovine testis: Purification, characterization and comparison with the enzymes from other tissues.

Cited by 11 publications

References 0 publications

Proline-specific endopeptidases from microbial sources: isolation of an enzyme from a Xanthomonas sp

Proline-specific endopeptidases from microbial sources: isolation of an enzyme from a Xanthomonas sp

Identification by ¹⁹F NMR of Traditional Chinese Medicinal Plants Possessing Prolyl Oligopeptidase Inhibitory Activity

Localization of the mRNA encoding prolyl endopeptidase in the rat brain and pituitary

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Prolyl endopeptidase from bovine testis: Purification, characterization and comparison with the enzymes from other tissues.

Cited by 11 publications

References 0 publications

Proline-specific endopeptidases from microbial sources: isolation of an enzyme from a Xanthomonas sp

Proline-specific endopeptidases from microbial sources: isolation of an enzyme from a Xanthomonas sp

Identification by 19F NMR of Traditional Chinese Medicinal Plants Possessing Prolyl Oligopeptidase Inhibitory Activity

Localization of the mRNA encoding prolyl endopeptidase in the rat brain and pituitary

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Identification by ¹⁹F NMR of Traditional Chinese Medicinal Plants Possessing Prolyl Oligopeptidase Inhibitory Activity