Prolidase Activity Research Articles

Structural basis for prolidase deficiency disease mechanisms.

All refined structure coordinates as well as the corresponding structure factor amplitudes have been deposited in the PDB under the accession numbers 5MBY, 5MBZ, 5MC0, 5MC1, 5MC2, 5MC3, 5MC4, 5MC5, 6H2P, 6H2Q.

Constituents of Propolis: Chrysin, Caffeic Acid, p-Coumaric Acid, and Ferulic Acid Induce PRODH/POX-Dependent Apoptosis in Human Tongue Squamous Cell Carcinoma Cell (CAL-27).

Propolis evokes several therapeutic properties, including anticancer activity. These activities are attributed to the action of polyphenols. Previously it has been demonstrated, that one of the most abundant polyphenolic compounds in ethanolic extracts of propolis are chrysin, caffeic acid, p-coumaric acid, and ferulic acid. Although their pro-apoptotic activity on human tongue squamous cell carcinoma cells (CAL-27) was established previously, the detailed mechanism of this process remains unclear. Considering the crucial role of proline metabolism and proline dehydrogenase/proline oxidase (PRODH/POX) in the regulation of cancer cell survival/apoptosis, we studied these processes in polyphenol-treated CAL-27 cells. All studied polyphenols evoked anti-proliferative activity, accompanied by increased PRODH/POX, P53, active caspases-3 and -9 expressions and decreased collagen biosynthesis, prolidase activity and proline concentration in CAL-27 cells. These data suggest that polyphenols of propolis induce PRODH/POX-dependent apoptosis through up-regulation of mitochondrial proline degradation and down-regulation of proline utilization for collagen biosynthesis.

Proline oxidase silencing induces proline-dependent pro-survival pathways in MCF-7 cells.

Proline degradation by proline dehydrogenase/proline oxidase (PRODH/POX) contributes to apoptosis or autophagy. The identification of specific pathway of apoptosis/survival regulation is the aim of this study. We generated knocked-down PRODH/POX MCF-7 breast cancer cells (MCF-7shPRODH/POX). PRODH/POX silencing did not affect cell viability. However, it contributed to decrease in DNA and collagen biosynthesis, increase in prolidase activity and intracellular proline concentration as well as increase in the expression of iNOS, NF-κB, mTOR, HIF-1α, COX-2, AMPK, Atg7 and Beclin-1 in MCF-7shPRODH/POX cells. In these cells, glycyl-proline (GlyPro, substrate for prolidase) further inhibited DNA and collagen biosynthesis, maintained high prolidase activity, intracellular concentration of proline and up-regulated HIF-1α, AMPK, Atg7 and Beclin-1, compared to GlyPro-treated MCF-7 cells. In MCF-7 cells, GlyPro increased collagen biosynthesis, concentration of proline and expression of caspase-3, cleaved caspases -3 and -9, iNOS, NF-κB, COX-2 and AMPKβ. PRODH/POX knock-down contributed to pro-survival autophagy pathways in MCF-7 cells and GlyPro-derived proline augmented this process. However, GlyPro induced apoptosis in PRODH/POX-expressing MCF-7 cells as detected by up-regulation of active caspases -3 and -9. The data suggest that PRODH/POX silencing induces autophagy in MCF-7 cells and GlyPro-derived proline supports this process.

Dexmedetomidine and Magnesium Sulfate: A Good Combination Treatment for Acute Lung Injury?

ABSTRACTObjectives: In this study, we aimed to investigate the therapeutic effects of magnesium sulfate (MgSO4) and dexmedetomidine (dex) in a model of acute lung injury (ALI). We determined whether concomitant administration decreased the inflammatory effects of hydrochloric acid (HCl)-induced ALI in a synergistic manner. Materials and Methods: In this study, 42 Sprague–Dawley rats were randomized into six groups: Group S (saline), Group SV (saline + mechanical ventilation), Group HCl (HCl), Group Dex (Dex), Group Mag (MgSO4), and Group DM (Dex + MgSO4). All groups except Group S were mechanically ventilated prior to HCl-induced ALI. Saline or HCl was administered via tracheostomy. Prior to treatment, HCl was administered to Group HCl, Group Dex, Group Mag, and Group DM to induce ALI. Dex and MgSO4 were administered intraperitoneally. The rats were monitored for 4 h after treatment to measure oxidative stress parameters in blood, and prolidase enzyme activity. Lung tissue damage were determined via histopathology. Results: A significant increase in heart rate and rapid desaturation was observed in HCl-administered groups. Treatment administration decreased the pulse values. Increased saturation values and decreased oxidative stress indices were observed in groups that were subsequently administered​ Dex and MgSO4. Serum prolidase activity increased significantly in Group HCl. Severe pathological findings were detected following HCl-induced ALI. Group Mag showed greater improvement in the pathology of HCl-induced ALI than did Group Dex. Administration of both Dex and MgSO4 did not improve the pathological scores. Conclusions: The antioxidant and anti-inflammatory effects of Dex and MgSO4 ameliorated the detrimental effects of HCI-induced ALI. However, adverse effects on hemodynamics and lung damage were observed when the two drugs were administered together.

Evaluation of Prolidase Activity, Oxidative Stress, and Antioxidant Enzyme Levels in Testicular and Penile Tissues after Human Chorionic Gonadotropin Treatment in Rats by Predicting Infertility and Erectile Dysfunction

Objectives: Prolidase plays a vital role in collagen turnover, matrix remodeling, and cell growth. We aimed to evaluate the association between treatment with chorionic gonadotropin and infertility and erectile dysfunction by investigating tissue prolidase activity, oxidative stress, and levels of antioxidant enzymes. Materials and Methods: The 16 male Wistar albino rats used in this study were randomly divided into 2 groups: rats treated with human chorionic gonadotropin (hCG) and control rats (n = 8 in each group). The rats in the hCG group were subcutaneously injected with 50 IU hCG daily for 15 days, while the rats in the control group were subcutaneously injected isotonic saline. All of the rats were sacrificed by a lethal overdose of sodium pentobarbital at the first month after hCG administration. Prolidase activity and levels of malonyl aldehyde, glutathione reductase, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) were estimated in the testicular and penile tissue. The testicles and penis were transversely dissected and placed in formalin. Results: Levels of prolidase and malonyl aldehyde in the testicular and penile tissues were significantly higher in the hCG group than in the control group (p < 0.001), while levels of glutathione reductase, SOD, GSH-Px, and CAT were significantly lower in the hCG group than in the control group (p < 0.001). Conclusions: In this study, we observed that treatment with hCG increased prolidase activity and oxidative stress and decreased the antioxidant capacity of penile and testicular tissues; therefore, this may affect fertility and erectile function.

Prolidase Activity and Oxidative Stress in Patients with Glioma

Prolidase Activity and Oxidative Stress in Patients with Glioma

Comparison of paraoxonase, prolidase activities and Hs-CRP, BDNF levels in patients with autogenous and reactive subtypes of obsessive compulsive disorder

Objective: Obsessions have been categorized as autogenous obsessions and reactive obsessions on the original of the cognitive theory and this study aimed to evaluate whether differences are found between obsessive compulsive disorder (OCD) subgroups in terms of biochemical markers. Methods: Thirty patients with obsessions defined as autogenous, 30 patients with obsessions defined as reactive group and 30 healthy volunteers were consisted the sample of the study. Sociodemographic Data Form, Yale-Brown Obsessive Compulsive Scale (Y-BOCS), Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI) were filled to the participants. The activity of paraoxanase, prolidase and Hs-CRP and brain-derived neurotrophic factor (BDNF) levels were detected. Results: There was no any statistically significant difference between autogenous and reactive obsession groups in terms of total mean score of Y-BOCS, BDI and BAI. Paraoxanase activity did not statistically significant difference between all groups. Prolidase activity showed significantly higher in reactive group than autogenous obsession and control groups. Serum Hs-CRP levels were not statistically different among participants of autogenous group, reactive group and control groups. Serum BDNF levels were significantly lower in autogenous obsession group rather than reactive obsession and control groups. Conclusion: This finding indicates that neurobiological pro-cesses have important role in the pathophysiology of autogenous obsessions.

Association of Tissue Transglutaminase with Nerve Growth Factor, Prolidase Activity and Oxidative Stress in Celiac Patients

Association of Tissue Transglutaminase with Nerve Growth Factor, Prolidase Activity and Oxidative Stress in Celiac Patients

Serum prolidase enzyme activity as a diagnostic marker for acute ischemic stroke

Objective: We aimed to investigate whether Serum Prolidase Activity (SPA) levels could be used as a potential diagnostic and/or prognostic biomarker in Acute Ischemic Stroke (AIS) patients or not. Materials and Methods: SPA levels were prospectively evaluated in 37 patients aged between 20 and 85 y who were admitted within 24 h of the onset of AIS. The control group included 37 healthy volunteers of similar age without any disease. Results: In AIS patients, mean SPA was significantly higher compared to healthy controls (1331 ± 399 pg/ml vs. 1169 ± 221 pg/ml, respectively; p=0.035). SPA was not correlated with age, gender, hypertension, diabetes, total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, hemoglobin, c-reactive protein, or hemoglobin A1c levels (p>0.05 for all comparisons). However, patients with new diagnostic atrial fibrillation had higher levels of prolidase acitvity than the others (1647 ± 403 pg/ml vs. 1270 ± 384 pg/ml, p=0.032). SPA levels were also uncorrelated with National Institutes of Health Stroke Scale, infarct volume, Trial of Org 10172 and the Oxfordshire Community Stroke Project classifications, and duration of hospitalization (p>0.05, for all comparisons). Conclusions: Increased levels of serum prolidase enzyme activity may be an independent predictor of AIS and may contribute to stroke pathophysiology. However, further studies with larger populations are needed to reveal the role of SPA in AIS.

Evaluation of serum prolidase activity and oxidative stress markers in men with BPH and prostate cancer

BackgroundProstate cancer (PCa) and benign prostatic hyperplasia (BPH) are diseases of elderly men and are related to increased oxidative stress (OS). Although prolidase has a role in collagen metabolism, it is also used to evaluate OS in many diseases. However, there is a lack of data about serum prolidase activity (SPA) in prostate cancer. The aim of this study was to evaluate and compare SPA levels in males with BPH and PCa.MethodsEvaluation was made of a total of 81 men who underwent transrectal ultrasound guided prostate biopsy for a definitive diagnosis due to high PSA levels. Patients were separated into 2 groups as BPH and PCa patients. Pre-biopsy malondialdehyde (MDA), superoxide dismutase (SOD), PSA levels and serum prolidase activities (SPA) were compared between the groups and the correlations of SPA with the other parameters were also investigated in both groups.ResultsBPH was diagnosed in 51 patients and PCa in 30. The mean age of patients was similar in both groups as 63.25 ± 5.81 years in the BPH group 65.30 ± 7.35 years in the PCa group(p:0.081). The median MDA and SOD levels were insignificantly increased in the PCa patients. SPA values were similar in BPH and PCa patients. SPA did not correlate with age, PSA, MDA or SOD levels in either group.ConclusionsOur study results revealed that serum prolidase activity is similar in BPH and PCa cases and is not correlated with MDA, SOD or PSA levels.

Acetylenic derivative of betulin induces apoptosis in endometrial adenocarcinoma cell line

Since betulin (Bet) and its acetylenic derivative, 28-O-propynoylbetulin (proBet) were shown to induce apoptosis in several cancer cell lines, we studied the mechanism of this process in human endometrial adenocarcinoma cells (EA). Previous studies suggested that this group of compounds affect prolidase activity (proline releasing enzyme from imidodipeptides) and collagen biosynthesis (proline utilizing process) providing substrate (proline) for proline oxidase (POX) dependent apoptosis. Here we provide evidence that Bet and proBet exhibit prolidase-inducing activity in EA cell line. However, in contrast to Bet, proBet inhibited collagen biosynthesis, increased intracellular proline concentration and induced apoptosis in EA cells, as detected by caspase-3, and -9 expressions and annexin V staining. Although POX expression was not affected by both compounds, the process of apoptosis was accompanied by increase in cytoplasmic level of proline. The mechanism for proBet-induced prolidase activity was found at the level of β1 integrin signaling. The inhibition of collagen biosynthesis was due to up-regulation of NF-κB p65, an inhibitor of collagen type I gene transcription. Although Bet and proBet induced expression of pro-apoptotic p53 in EA cells, the effect of proBet on the processes was much stronger. In contrast to proBet, Bet strongly induced expression of pro-survival factors, HIF-1α and VEGF. The data suggest that massive production of proline by proBet-dependent activation of prolidase and inhibition of proline utilization for collagen biosynthesis may represent mechanism for POX-dependent apoptosis in EA cells.

Improved Hydrolysis of Organophosphorus Compounds by Engineered Human Prolidases.

Human prolidase has weak hydrolytic activity for toxic organophosphorus compounds including diisopropyl fluorophosphates (DFP), chemical warfare nerve agents and pesticides. In order to use human prolidase as a catalytic bioscavenger against toxic organophosphorus compound exposure, protein engineering is an important issue to improve the catalytic activity of human prolidase towards the hydrolysis of toxic organophosphorus compounds. We developed two human prolidase mutants, A252R and P365R, with a single amino acid substitution using in silico analysis based on the sequence, protein structure and stability to improve the catalytic activity of human prolidase towards DFP hydrolysis. Our results showed that the catalytic efficiencies of A252R and P365R towards DFP hydrolysis were 1.23- and 1.36-fold increases, respectively, than that of the wild type, while the prolidase activities of A252R and P365R towards Leu-Pro hydrolysis were 0.88- and 0.78-fold decreases that of the wild type, respectively, indicating that substitution mutations of A252R and P365R in human prolidase show improved hydrolytic activity for toxic organophosphorus compounds. We report here that by introducing either the A252R or P365R substitution mutation, the structural changes affecting catalytic turnover rate and substrate binding affinity are valuable in improving the catalytic activity of human prolidase towards toxic organophosphorus compound hydrolysis.

Serum prolidase enzyme activity in obese subjects and its relationship with oxidative stress markers

BackgroundThe relationship between increased serum enzyme activity of prolidase and increased rate of collagen turnover in the arterial wall has been asserted in previous studies. Collagen reflects much of the strength to the connective tissue involved in the arterial wall. Atherosclerosis is very common vessel disease and oxidative stress plays a pivotal role in the etiopathogenesis. Our objective was to examine the serum enzyme activity of prolidase and its possible relationships with oxidative stress parameters in obese subjects. MethodsOur present study was conducted 27 obese subjects and 26 age-matched healthy control subjects. The serum enzyme activity of prolidase in all study population was evaluated spectrophotometrically. Oxidative stress levels in obese subjects were analyzed with total antioxidant capacity (TAC) and total oxidant status (TOS) as well as oxidative stress index (OSI). ResultsObese subjects have higher serum TOS and OSI indicators as well as prolidase activity than those in control subjects (for all; p<0.001). Moreover, obese subjects have lower levels of TAC than in those in healthy subjects (p<0.001). In the Pearson's correlation analysis, enzyme activity of prolidase was positively related with TOS (p<0.001, r=0.529) and OSI (p<0.001, r=0.519) as well as BMI (p<0.001, r=0.692) and inversely related with TAC (p<0.05, r=−0.405) in obese subjects. ConclusionsIncreased serum prolidase activity and decreased antioxidant levels are likely to be a results of increased of oxidative stress levels in obese subjects. The significantly correlation between increased oxidative stress and increased prolidase activity may play a pivotal role in etiopathogenesis of atherosclerotic cardiovascular diseases in obese subjects.

Substrate specificity and reaction mechanism of human prolidase

The refined structure coordinates as well as the corresponding structure factor amplitudes have been deposited in the PDB under the accession numbers 5M4G, 5M4J, 5M4L, and 5M4Q.

Circulating Prolidase Activity in Patients with Myocardial Infarction.

Collagen is a major determinant of atherosclerotic plaque stability. Thus, identification of differences in enzymes that regulate collagen integrity could be useful for predicting susceptibility to atherothrombosis or for diagnosing plaque rupture. In this study, we sought to determine whether prolidase, the rate-limiting enzyme of collagen turnover, differs in human subjects with acute myocardial infarction (MI) versus those with stable coronary artery disease (CAD). We measured serum prolidase activity in 15 patients with stable CAD and 49 patients with acute MI, of which a subset had clearly defined thrombotic MI (n = 22) or non-thrombotic MI (n = 12). Prolidase activity was compared across study time points (at cardiac catheterization, T0; 6 h after presentation, T6; and at a quiescent follow-up, Tf/u) in acute MI and stable CAD subjects. We performed subgroup analyses to evaluate prolidase activity in subjects presenting with acute thrombotic versus non-thrombotic MI. Although prolidase activity was lower at T0 and T6 versus the quiescent phase in acute MI and stable CAD subjects (p < 0.0001), it was not significantly different between acute MI and stable CAD subjects at any time point (T0, T6, and Tf/u) or between thrombotic and non-thrombotic MI groups. Preliminary data from stratified analyses of a small number of diabetic subjects (n = 8) suggested lower prolidase activity in diabetic acute MI subjects compared with non-diabetic acute MI subjects (p = 0.02). Circulating prolidase is not significantly different between patients with acute MI and stable CAD or between patients with thrombotic and non-thrombotic MI. Further studies are required to determine if diabetes significantly affects prolidase activity and how this might relate to the risk of MI.

Differential effect of platelet-rich plasma fractions on β1-integrin signaling, collagen biosynthesis, and prolidase activity in human skin fibroblasts.

The study was conducted to evaluate the effects of platelet-rich plasma (PRP), supernatant of PRP (SPRP) obtained by centrifugation, and supernatant of activated PRP (SActi-PRP) obtained by Ca2+ solution-treated PRP on collagen biosynthesis, prolidase activity, and β1-integrin signaling in cultured human skin fibroblasts. Incubation of fibroblasts with 5% PRP for 24 h contributed to ~5-fold increase in collagen biosynthesis compared to the control. In the cells treated with 5% of SPRP or SActi-PRP, collagen biosynthesis showed a 3-fold increase of the control. PRP, SPRP, and SActi-PRP stimulated prolidase activity similar to collagen biosynthesis. Collagen biosynthesis and prolidase activity are regulated by β1-integrin receptor signaling. Incubation of fibroblasts with PRP for 24 h contributed to a dose-dependent increase in the expression of β1-integrin receptor, while SActi-PRP increased the process to a much lower extent. SPRP had no effect on the β1-integrin receptor expression. All the studied fractions of blood increased the expression of FAK as well as the expression of phosphorylated MAP-kinases. However, PRP was found to be the most effective stimulator of expression of these particular kinases. These studies suggest that a complex of factors, including growth factors, adhesion molecules, and prolidase contained in PRP, all evoke growth and collagen-promoting activities in human dermal fibroblasts.

Serum prolidase activity in systemic sclerosis.

Systemic sclerosis, also known as scleroderma, is a complex systemic inflammatory autoimmune disease that targets the vasculature and connective tissue-producing cells and components of the innate and adaptive immune systems. The disease is characterized by a hardening of the skin and an increased synthesis of collagen . Prolidase is a specific imidodipeptidase involved in collagen degradation. The aim of this study was to search the serum prolidase activity (SPA) in the two subtypes of systemic sclerosis: diffuse and limited cutaneous systemic sclerosis. For this purpose, 35 patients diagnosed with systemic sclerosis (24 diffuse and 11 limited) and 41 healthy control subjects were included in the study. SPA was determined using Myara's method, which is a modification of Chinard's method. SPA did not differ between the scleroderma patients and controls (p=0.467). However, SPA was significantly lower in diffuse form than in both limited form and control subjects (p=0.021 and p=0.024, respectively). SPA also did not differ between the limited form and control subjects (p=0.145). Scleroderma is characterized by excessive deposition of collagen and tissue fibrosis due to the reduced collagen degradation. SPA is reduced in scleroderma patients, especially in diffuse form. Circulating autoantibodies, oxidative stress, and decreased physical activity may contribute to this process.

Exogenous proline stimulates type I collagen and HIF-1\u03b1 expression and the process is attenuated by glutamine in human skin fibroblasts

Abundance of proline (Pro) in collagen molecule led us to investigate whether Pro supply affects collagen biosynthesis in human skin fibroblasts. Treatment of the cells with milimolar concentrations (5 and 10 mM) of Pro for 24 and 48 h contributed to increase in α1 subunit of collagen type I (COL1A1) expression in both cells and culture medium. However, the effect was more pronounced in glutamine-free medium. In such condition, Pro induced collagen expression by about twofold in the cells, while in the medium only by about 30% during 24 h incubation, compared to control. In the presence of glutamine (Gln), exogenous Pro stimulated intracellular collagen expression only by about 30% during 24 h of fibroblasts incubation, and it was not accompanied by adequate increase of collagen secretion into medium. Gln alone stimulated the processes by about 2–3 fold during the course of the experiment. Pro-dependent increase in collagen expression in Gln-free medium was accompanied by increase in prolidase activity and expression of pAkt. In both Gln-free medium and Gln-supplemented medium, Pro induced expression of p53 and HIF-1α. The data suggest that availability of Gln, as a substrate for Pro biosynthesis, determine the utilization of exogenous Pro for the collagen biosynthesis.

Comparison of protective effect of ascorbic acid on redox and endocannabinoid systems interactions in in vitro cultured human skin fibroblasts exposed to UV radiation and hydrogen peroxide

The mechanisms of biological activity of commonly used natural compounds are constantly examined. Therefore, the aim of this study was to compare ascorbic acid efficacy in counteracting the consequences of UV and hydrogen peroxide treatment on lipid mediators and their regulative action on antioxidant abilities. Skin fibroblasts exposed to UVA and UVB irradiation, treated with hydrogen peroxide and ascorbic acid. The redox system was estimated through reactive oxygen species (ROS) generation (electron spin resonance spectrometer) and antioxidants level/activity (HPLC/spectrometry) which activity was evaluated by the level of phospholipid metabolites: 4-hydroxynonenal, malondialdehyde, 8-isoprostanes and endocannabinoids (GC/LC-MS) in the human skin fibroblasts. Protein and DNA oxidative modifications were also determined (LC). The expression of nuclear factor erythroid 2-related factor 2 (Nrf2), its activators and inhibitors as well as pro/anti-apoptotic proteins and endocannabinoid receptors was examined (Western blot) and collagen metabolism was evaluated by collagen biosynthesis and prolidase activity (spectrometry). UVA and UVB irradiation and hydrogen peroxide treatment enhanced activity of xanthine and NADPH oxidases resulting in ROS generation as well as diminution of antioxidant phospholipid protection (glutathione peroxidase-glutathione-vitamin E), what led to increased lipid peroxidation and decreased endocannabinoids level. Dysregulation of cannabinoid receptors expression and environment of transcription factor Nrf2 caused apoptosis induction. Ascorbic acid partially prevented ROS generation, antioxidant capacity diminution and endocannabinoid systems disturbances but only slightly protected macromolecules such as phospholipid, protein and DNA against oxidative modifications. However, ascorbic acid significantly prevented decrease in collagen type I biosynthesis. Ascorbic acid in similar degree prevents UV (UVA and UVB) and hydrogen peroxide-dependent redox imbalance. However, this antioxidant cannot efficiently protect cellular macromolecules and avert metabolic dysregulation leading to apoptosis.

SERUM PROLIDASE ACTIVITY IN STABLE CORONARY ARTERY DISEASE AND ACUTE MYOCARDIAL INFARCTION

Background: Collagen is a major determinant of atherosclerotic plaque stability and prolidase is a rate-limiting enzyme in the breakdown and formation of collagen. We sought to determine prolidase activity at the time of acute myocardial infarction (MI) as compared to quiescent state and to patients