Characterization of fibroblast-derived prolidase

Oono, Takashi; Yasutomi, Hiroshi; Ohhashi, Toshitaka; Kodama, Hiroyuki; Arata, Jirô

doi:10.1016/0923-1811(90)90588-5

Cited by 15 publications

(9 citation statements)

References 10 publications

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“…The substrate specificity of both tagged and untagged enzymes obtained from eukaryotic and prokaryotic sources was indistinguishable from that of endogenous fibroblast prolidase, the preferred substrate being Gly-Pro, followed by Ala-Pro. This is in accordance with data previously reported for human endogenous prolidase from fibroblasts and erythrocytes [33][34][35][36][37][38][39][40].…”

Section: Discussionsupporting

confidence: 93%

Human recombinant prolidase from eukaryotic and prokaryotic sources

Lupi¹,

Torre²,

Campari

et al. 2006

The FEBS Journal

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Prolidase is a Mn2+‐dependent dipeptidase that cleaves imidodipeptides containing C‐terminal proline or hydroxyproline. In humans, a lack of prolidase activity causes prolidase deficiency, a rare autosomal recessive disease, characterized by a wide range of clinical outcomes, including severe skin lesions, mental retardation, and infections of the respiratory tract. In this study, recombinant prolidase was produced as a fusion protein with an N‐terminal histidine tag in eukaryotic and prokaryotic hosts and purified in a single step using immobilized metal affinity chromatography. The enzyme was characterized in terms of activity against different substrates, in the presence of various bivalent ions, in the presence of the strong inhibitor Cbz‐Pro, and at different temperatures and pHs. The recombinant enzyme with and without a tag showed properties mainly indistinguishable from those of the native prolidase from fibroblast lysate. The protein yield was higher from the prokaryotic source, and a detailed long‐term stability study of this enzyme at 37 °C was therefore undertaken. For this analysis, an ‘on‐column’ digestion of the N‐terminal His tag by Factor Xa was performed. A positive effect of Mn2+ and GSH in the incubation mixture and high stability of the untagged enzyme are reported. Poly(ethylene glycol) and glycerol had a stabilizing effect, the latter being the more effective. In addition, no significant degradation was detected after up to 6 days of incubation with cellular lysate. Generation of the prolidase in Escherichia coli, because of its high yield, stability, and similarity to native prolidase, appears to be the best approach for future structural studies and enzyme replacement therapy.

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Section: Discussionsupporting

confidence: 93%

Human recombinant prolidase from eukaryotic and prokaryotic sources

Lupi¹,

Torre²,

Campari

et al. 2006

The FEBS Journal

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“…Prolidase I has a relative molecular mass of about 112000 and the highest activity towards GlyPro, whereas prolidase II has a relative molecular mass of about 185000 and is mainly active towards Met-Pro. Characteristics of the isozymes in patients and controls have been discussed in detail (12)(13)(14)(15)(16). The present paper also reports the prolidase activities for X-Hyp in patients, in their mother and in controls, which are closely correlated with the quantities of XHyp excreted in the urine.…”

Section: Introductionmentioning

confidence: 81%

The Use of Liquid Chromatography-Mass Spectrometry for the Identification and Quantification of Urinary Iminodipeptides in Prolidase Deficiency

Sugahara

Ohno²,

Arata³

et al. 1993

Clinical Chemistry and Laboratory Medicine

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It has been reported that the urine of patients with prolidase deficiency contains various iminodipeptides with a carboxyl-terminal proline (hydroxyproline). These iminodipeptides have hitherto been detected indirectly by acid hydrolysis or enzymatic digestion, followed by amino acid analysis. In the present study, it was shown that X-Pro could be distinguished from Pro-X when the iminodipeptides were analysed directly by liquid chromatography coupled with atmospheric pressure ionization mass Spectrometry (LC/API-MS), with scanning of the protonated molecule ions ([M + H]+). The same procedure also successfully quantified urinary iminodipeptides from patients with prolidase deficiency. A quantitative investigation of two siblings with prolidase deficiency revealed that the patient with severe clinical symptoms excreted more iminodipeptides than the other who did not have serious symptoms. LC/API-MS also revealed iminodipeptides (Gly-Hyp and Pro^Hyp) in the urine of the mother of the patients and in normal volunteers. Patients excreted much more Pro-Hyp than normal volunteers, whereas no quantitative differences were found between the mother and controls. In patients, the excretion of large quantities of X-Pro is due to their very low prolidase activity towards this type of substrate. In the erythrocytes of patients, prolidase activity towards X-Hyp was extremely low; even in the mother and normal volunteers, it was remarkably low in comparison with the activity against X-Pro.

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“…One of the consequences of this kind of inflammatory diseases is dysregulation in the metabolism of collagen and its interaction with cell surface integrin receptors. Although extracellular metalloproteinases initiate the breakdown of collagen in tissues, the final step of its degradation is mediated by prolidase (1)(2)(3)(4).…”

Section: Introductionmentioning

confidence: 99%

“…3.4.13.9) is a cytosolic Mn(II)activated metalloproteinase that specifically hydrolyzes imidodipeptides and imidotripeptides with C-terminal proline or hydroxyproline, releases these two amino acids for collagen re-synthesis and cell growth. This enzyme has two forms such as prolidase I (M W : 105,000) and prolidase II (M W : 151,000) but only prolidase I has been found in human plasma (2,3). Prolidase deficiency is a rare autosomal recessive disease characterized by chronic ulcerative dermatitis, splenomegaly, mental retardation, frequent infections, and massive urinary excretion of iminodipeptides (4,5).…”

Section: Introductionmentioning

confidence: 99%

Prolidase activity dysregulation and its correlation with oxidative-antioxidative status in chronic obstructive pulmonary disease

Gençer

Aksoy

Dagli

et al. 2011

J. Clin. Lab. Anal.

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Background: Chronic obstructive pulmonary disease (COPD) is a consequence of an underlying chronic inflammatory disorder of the airways that is usually progressive and causes dysregulation in the metabolism of collagen. Prolidase has an important role in the recycling of proline for collagen synthesis and cell growth. Objective: We measured and compared prolidase activity in healthy individuals with COPD patients to find out that whether its activity might reflect disturbances of collagen metabolism in the patients. We also investigated oxidative-antioxidative status and its relationship with prolidase activity in this disease. Methods: Thirty voluntary patients with COPD and 30 healthy control subjects with similar age range and sex were included into the study. Plasma prolidase activities, total antioxidant capacity (TAC) and lipid peroxidation (LPO) levels were measured in the patient and control groups.Results: Plasma prolidase activity and TAC levels were significantly lower, and LPO levels were significantly higher in the patients than those in the control subjects (Po0.05, Po0.001, and Po0.001, respectively). Significant correlations were detected between plasma prolidase activity and TAC and LPO levels in the patients group (r 5 0.679, Po0.001; r 5 À426, Po0.05, respectively). Conclusions:The results suggest that oxidative-antioxidative balance and collagen turnover are altered by the development of COPD in human lungs, and prolidase activity may reflect disturbances of collagen metabolism in this pulmonary disease. Monitoring of plasma prolidase activity and oxidativeantioxidative balance may be useful in evaluating fibrotic processes and oxidative damage in the chronic inflammatory lung disease in human.

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Characterization of fibroblast-derived prolidase

Cited by 15 publications

References 10 publications

Human recombinant prolidase from eukaryotic and prokaryotic sources

Human recombinant prolidase from eukaryotic and prokaryotic sources

The Use of Liquid Chromatography-Mass Spectrometry for the Identification and Quantification of Urinary Iminodipeptides in Prolidase Deficiency

Prolidase activity dysregulation and its correlation with oxidative-antioxidative status in chronic obstructive pulmonary disease

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