Pyrrolidonyl Peptidase in Animal, Plant and Human Tissues. Occurrence and Some Properties of the Enzyme

Szewczuk, A; Kwiatkowska, J

doi:10.1111/j.1432-1033.1970.tb00980.x

Cited by 94 publications

(30 citation statements)

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“…Enzymes of the second type are present both in mammalian tissues (40) and in various bacterial species (30). They show a general specificity for all N-terminal pyroglutamyl residues and are highly sensitive to thiol-blocking agents (4,24,(39)(40)(41)43). The enzyme described in this article belongs to the second class, like all other bacterial Pcps characterized to date, and shows strong homology to sequenced Pcps from S. pyogenes, B. subtilis, and B. amyloliquefaciens.…”

Section: Discussionmentioning

confidence: 87%

“…Enzymes of the first class are found in mammalian tissues; they are specific for the thyrotropinreleasing hormone peptide (31,32) and sensitive to metal chelators but insensitive to thiol-blocking agents (13). Enzymes of the second type are present both in mammalian tissues (40) and in various bacterial species (30). They show a general specificity for all N-terminal pyroglutamyl residues and are highly sensitive to thiol-blocking agents (4,24,(39)(40)(41)43).…”

Section: Discussionmentioning

confidence: 99%

“…The purified enzyme was assayed at various temperatures (25,30,35,40, and 45°C) after 10 min of incubation to determine the optimum reaction temperature.…”

Section: Methodsmentioning

confidence: 99%

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Characterization of the pcp gene of Pseudomonas fluorescens and of its product, pyrrolidone carboxyl peptidase (Pcp)

Gonzalès

Robert‐Baudouy

1994

J Bacteriol

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The gene pcp, encoding pyrrolidone carboxyl peptidase (Pcp), from Pseudomonasfluorescens MFO was cloned and its nucleotide sequence was determined. This sequence contains a unique open reading frame (pcp) coding for a polypeptide of 213 amino acids (Mr 22,441) which has significant homology to the Pcps from Streptococcus pyogenes, Bacillus subtilis, and Bacilus amyloliquefaciens. Comparison of the four Pcp sequences revealed two highly conserved motifs which may be involved in the active site of these enzymes. The cloned Pcp from P. fluorescens was purified to homogeneity and appears to exist as a dimer. This enzyme displays a Michaelis constant of 0.21 mM with L-pyroglutamyl-,1-naphthylamide as the substrate and an absolute substrate specificity towards N-terminal pyroglutamyl residues. Studies of inhibition by chemical compounds revealed that the cysteine and histidine residues are essential for enzyme activity. From their conservation in the four enzyme sequences, the Cys-144 and His-166 amino acids are proposed to form a part of the active site of these enzymes.Pyrrolidone carboxyl peptidases (Pcps) (EC 3.4.11.8) are aminopeptidases that selectively remove amino-terminal Lpyroglutamic acid (Pyr) from peptides and proteins (15). These enzymes are present in many bacteria (30) and also in plant, animal, and human tissues (40).Preliminary studies of Pcps have consisted of purification of these enzymes from different bacterial strains (1,24,39,41,43) with a view to using them in protein sequencing to unblock proteins and polypeptides with an N-terminal pyroglutamyl residue prior to sequential Edman degradation (16). These bacterial Pcps, which are cysteine peptidases, showed an absolute specificity for pyroglutamyl residues located at the N terminus of peptides (41, 43) and for chromogenic substrates, such as L-pyroglutamyl-,B-naphthylamide (Pyr-3NA) (41), pyroglutamyl-p-nitroanilide, and pyroglutamyl-4-methylcoumarinylamide (17).Mammalian Pcps were later analyzed and shown to be involved in hormonal and neuronal metabolism (6,8,44). Two different types of Pcps have been detected in mammals. The first, which is a metallopeptidase, is specific for the pyroglutamyl-neuropeptide thyrotropin-releasing hormone (Pyr-HisPro-NH2) and is responsible for regulating the concentration of this neuropeptide (13,31,32). The second is a cysteine peptidase with a general specificity for N-terminal pyroglutamyl residues (7, 10). This Pcp, the function of which has not yet been determined, seems to be more closely related to bacterial Pcps than the first with respect to size, substrate specificity, and sensitivity to inhibitors.Recently, the structural genes encoding Pcps from the gram-positive bacteria Streptococcus pyogenes (12), Bacillus subtilis (3), and Bacillus amyloliquefaciens (45) have been cloned and characterized. The three deduced amino acid sequences are strongly homologous and display long conserved domains. The absence of homology with other peptidases suggests that these Pcps belong to a new class of peptidases * ...

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

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Characterization of the pcp gene of Pseudomonas fluorescens and of its product, pyrrolidone carboxyl peptidase (Pcp)

Gonzalès

Robert‐Baudouy

1994

J Bacteriol

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“…The pyrrolidone carboxyl peptidases (Pcps) are a group of exopeptidases responsible for the removal of N-terminal pyroglutamate residues from a variety of peptides and proteins (Doolittle and Armentrout 1968). This enzyme was found to be widely distributed (Szewczuk and Kwiatkowska 1970), but its physiological role is still not clear. Pcp activity in archaea and eubacteria is thought to be involved in detoxification processes and nutrient metabolism (Awadé et al 1994).…”

Section: Pyrrolidone-carboxylate Peptidasementioning

confidence: 99%

Down-regulated gene expression of cold acclimated Poncirus trifoliata

Zhang¹,

Lang²,

Ebel³

et al. 2005

Can. J. Plant Sci.

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. 2005. Down-regulated gene expression of cold acclimated Poncirus trifoliata. Can. J. Plant Sci. 85: 417-424. Citrus sp. are important commercial fruit crops throughout the world that are occasionally devastated by subfreezing temperatures. Poncirus trifoliata (maximum freeze tolerance of -26°C) is a close relative of commercial Citrus sp. (maximum freeze tolerance of -10°C) that has been used in breeding programs to develop more cold-hardy genotypes and as a rootstock to enhance freeze tolerance of the scion. Species with greater freeze tolerance vary in gene expression during cold acclimating temperatures. mRNA differential display (DDRT-PCR) and quantitative relative RT-PCR were used to study down regulation of gene expression in intact P. trifoliata exposed to a gradual cold acclimation regime to enhance our understanding of the mechanism that makes this specie so freeze tolerant. Six down-regulated genes were isolated and sequenced. These down-regulated genes showed high homology to the following known genes: chlorophyll a/b binding protein, photosystem II OEC 23, carbonic anhydrase, tumor related protein, pyrrolidone-carboxylate peptidase and β-galactosidase. Photoprotection and the global control of gene expression related to photosynthesis appear to be important mechanisms for cold acclimation of P. trifoliata. Poncirus trifoliata (tolérance maximale au gel de -26°C) est un proche parent du genre Citrus (tolérance maximale au gel de -10°C) employé dans les programmes d'hybridation pour produire des génotypes plus rustiques et des porte-greffe qui accroîtront la tolérance au gel du greffon. L'expression des gènes des espèces qui tolèrent bien le froid varie lors des températures conduisant à l'acclimatation. Les auteurs ont recouru au criblage différentiel de l'ARNm (DDRT-PCR) et à la PCR-CDNA quantitative relative pour étudier la régulation en aval de l'expression des gènes chez les plants de P. trifoliata restés intacts après une acclimatation progressive au froid afin de nous aider à mieux comprendre le mécanisme par lequel cette espèce réussit à survivre si bien au gel. Ils ont ainsi identifié et séquencé six gènes régulés en aval. Ces gènes se caractérisent par une grande homologie avec les gènes connus suivants : protéine a/b de liaison à la chlorophylle, photosystème II OEC 23, anhydrase carbonique, protéine associée aux tumeurs, pyrrolidone-carboxylate peptidase et β-galactosidase. La photoprotection et le contrôle général de l'expression des gènes associés à la photosynthèse semblent jouer un rôle important dans l'adaptation de P. trifoliata au froid.

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“…The Pcp activity was first reported three decades ago (12) and has since been demonstrated in bacteria, plants, and animal and human tissues (33). Preliminary studies of Pcps have concentrated on their purification from bacterial strains in order to investigate the biochemical properties of these unusual enzymes (16,32,34).…”

mentioning

confidence: 99%

Mutational analysis of the active site of Pseudomonas fluorescens pyrrolidone carboxyl peptidase

Saux¹,

Gonzales²,

Robert‐Baudouy³

1996

J Bacteriol

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On the basis of chemical inhibition studies and a multiple alignment of four pyrrolidone carboxyl peptidase (Pcp) amino acid sequences, seven conserved residues of the Pseudomonas fluorescens Pcp, which might be important for enzyme activity, have been modified by site-directed mutagenesis experiments. Wild-type and mutant Pcps were expressed in Escherichia coli, purified, and characterized by the ability to cleave the synthetic chromogenic substrate pyroglutamyl-␤-naphthylamide and the dipeptide pyroglutamyl-alanine. Substitution of Glu-10 and Glu-22 by Gln led to enzymes which displayed catalytic properties and sensitivities to 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide similar to those of the wild-type Pcp. These residues are not essential for the catalytic activity. Replacement of Asp-89 by Asn and Ala resulted in enzymes which retained nearly 25% of activity and which had no activity, respectively. Substitution of the Cys-144 and His-166 residues by Ala and Ser, respectively, resulted in inactive enzymes. Proteins with changes of Glu-81 to Gln and Asp-94 to Asn were not detectable in crude extract and were probably unstable in bacteria. Our results are consistent with the proposal that Cys-144 and His-166 constitute the nucleophilic and imidazole residues of the Pcp active site, while residue Glu-81, Asp-89, or Asp-94 might constitute the third part of the active site. These results lead us to propose Pcps as a new class of thiol aminopeptidases.

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Pyrrolidonyl Peptidase in Animal, Plant and Human Tissues. Occurrence and Some Properties of the Enzyme

Cited by 94 publications

References 9 publications

Characterization of the pcp gene of Pseudomonas fluorescens and of its product, pyrrolidone carboxyl peptidase (Pcp)

Characterization of the pcp gene of Pseudomonas fluorescens and of its product, pyrrolidone carboxyl peptidase (Pcp)

Down-regulated gene expression of cold acclimated Poncirus trifoliata

Mutational analysis of the active site of Pseudomonas fluorescens pyrrolidone carboxyl peptidase

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