scholarly journals CLytA-DAAO Chimeric Enzyme Bound to Magnetic Nanoparticles. A New Therapeutical Approach for Cancer Patients?

2021 ◽  
Vol 22 (3) ◽  
pp. 1477
Author(s):  
María Fuentes-Baile ◽  
Elizabeth Pérez-Valenciano ◽  
Pilar García-Morales ◽  
Camino de Juan Romero ◽  
Daniel Bello-Gil ◽  
...  
Keyword(s):  
Magnetic Nanoparticles ◽  
Colorectal Carcinoma ◽  
Acid Oxidase ◽  
Oxygen Species ◽  
Cell Models ◽  
Amino Acid Oxidase ◽  
Direct Addition ◽  
Alginate Capsules ◽  
Anti Cancer ◽  
The Stability

D-amino acid oxidase (DAAO) is an enzyme that catalyzes the oxidation of D-amino acids generating H2O2. The enzymatic chimera formed by DAAO bound to the choline-binding domain of N-acetylmuramoyl-L-alanine amidase (CLytA) induces cytotoxicity in several pancreatic and colorectal carcinoma and glioblastoma cell models. In the current work, we determined whether the effect of CLytA-DAAO immobilized in magnetic nanoparticles, gold nanoparticles, and alginate capsules offered some advantages as compared to the free CLytA-DAAO. Results indicate that the immobilization of CLytA-DAAO in magnetic nanoparticles increases the stability of the enzyme, extending its time of action. Besides, we compared the effect induced by CLytA-DAAO with the direct addition of hydrogen peroxide, demonstrating that the progressive generation of reactive oxygen species by CLytA-DAAO is more effective in inducing cytotoxicity than the direct addition of H2O2. Furthermore, a pilot study has been initiated in biopsies obtained from pancreatic and colorectal carcinoma and glioblastoma patients to evaluate the expression of the main genes involved in resistance to CLytA-DAAO cytotoxicity. Based on our findings, we propose that CLytA-DAAO immobilized in magnetic nanoparticles could be effective in a high percentage of patients and, therefore, be used as an anti-cancer therapy for pancreatic and colorectal carcinoma and glioblastoma.

scholarly journals d-amino acid oxidase promotes cellular senescence via the production of reactive oxygen species

2019 ◽  
Vol 2 (1) ◽  
pp. e201800045 ◽  
Author(s):  
Taiki Nagano ◽  
Shunsuke Yamao ◽  
Anju Terachi ◽  
Hidetora Yarimizu ◽  
Haruki Itoh ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Amino Acids ◽  
Dna Damage ◽  
Amino Acid ◽  
Cellular Senescence ◽  
Reactive Oxygen ◽  
Acid Oxidase ◽  
Dependent Manner ◽  
Oxygen Species ◽  
Amino Acid Oxidase

d-amino acid oxidase (DAO) is a flavin adenine dinucleotide (FAD)–dependent oxidase metabolizing neutral and polard-amino acids. Unlikel-amino acids, the amounts ofd-amino acids in mammalian tissues are extremely low, and therefore, little has been investigated regarding the physiological role of DAO. We have recently identifiedDAOto be up-regulated in cellular senescence, a permanent cell cycle arrest induced by various stresses, such as persistent DNA damage and oxidative stress. Because DAO produces reactive oxygen species (ROS) as byproducts of substrate oxidation and the accumulation of ROS mediates the senescence induction, we explored the relationship between DAO and senescence. We found that inhibition of DAO impaired senescence induced by DNA damage, and ectopic expression of wild-type DAO, but not enzymatically inactive mutant, enhanced it in an ROS-dependent manner. Furthermore, addition ofd-amino acids and riboflavin, a metabolic precursor of FAD, to the medium potentiated the senescence-promoting effect of DAO. These results indicate that DAO promotes senescence through the enzymatic ROS generation, and its activity is regulated by the availability of its substrate and coenzyme.

scholarly journals CLytA-DAAO, Free and Immobilized in Magnetic Nanoparticles, Induces Cell Death in Human Cancer Cells

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 222 ◽  
Author(s):  
María Fuentes-Baile ◽  
Daniel Bello-Gil ◽  
Elizabeth Pérez-Valenciano ◽  
Jesús M. Sanz ◽  
Pilar García-Morales ◽  
...  
Keyword(s):  
Cell Death ◽  
Magnetic Nanoparticles ◽  
Tumor Cells ◽  
Cell Lines ◽  
Epigenetic Therapy ◽  
Acid Oxidase ◽  
Glioblastoma Cell ◽  
Amino Acid Oxidase ◽  
Apoptosis Pathway ◽  
Glioblastoma Cell Lines

D-amino acid oxidase (DAAO) catalyzes the oxidation of D-amino acids generating hydrogen peroxide, a potential producer of reactive oxygen species. In this study, we used a CLytA-DAAO chimera, both free and bound to magnetic nanoparticles, against colon carcinoma, pancreatic adenocarcinoma, and glioblastoma cell lines. We found that the enzyme induces cell death in most of the cell lines tested and its efficiency increases significantly when it is immobilized in nanoparticles. We also tested this enzyme therapy in non-tumor cells, and we found that there is not cell death induction, or it is significantly lower than in tumor cells. The mechanism triggering cell death is apparently a classical apoptosis pathway in the glioblastoma cell lines, while in colon and pancreatic carcinoma cell lines, CLytA-DAAO-induced cell death is a necrosis. Our results constitute a proof of concept that an enzymatic therapy, based on magnetic nanoparticles-delivering CLytA-DAAO, could constitute a useful therapy against cancer and besides it could be used as an enhancer of other treatments such as epigenetic therapy, radiotherapy, and treatments based on DNA repair.

Identification of a Novel Spinal Dorsal Horn Astroglial d-Amino Acid Oxidase–Hydrogen Peroxide Pathway Involved in Morphine Antinociceptive Tolerance

2014 ◽  
Vol 120 (4) ◽  
pp. 962-975 ◽  
Author(s):  
Nian Gong ◽  
Xin-Yan Li ◽  
Qi Xiao ◽  
Yong-Xiang Wang
Keyword(s):  
Hydrogen Peroxide ◽  
Morphine Tolerance ◽  
Acid Oxidase ◽  
Oxygen Species ◽  
Morphine Treatment ◽  
Amino Acid Oxidase ◽  
Chronic Morphine ◽  
Hot Plate ◽  
Antinociceptive Tolerance ◽  
Interfering Rna

Abstract Background: d-Amino acid oxidase (DAAO) is a flavin adenine dinucleotide-dependent peroxisomal flavoenzyme which is almost exclusively expressed within astrocytes in the spinal cord. DAAO catalyzes oxidation of d-amino acids to hydrogen peroxide, which is a stable and less active reactive oxygen species, and may represent a final form of reactive oxygen species. This study tested the hypothesis that the spinal astroglial DAAO–hydrogen peroxide pathway plays an important role in the development of morphine antinociceptive tolerance. Methods: Rat and mouse formalin, hot-plate, and tail-flick tests were used, and spinal DAAO expression and hydrogen peroxide level were measured. Sample size of animals was six in each study group. Results: Subcutaneous and intrathecal DAAO inhibitors, including 5-chloro-benzo[d]isoxazol-3-ol, AS057278, and sodium benzoate, completely prevented and reversed morphine antinociceptive tolerance in the formalin, hot-plate, and tail-immersion tests, with a positive correlation to their DAAO inhibitory activities. Intrathecal gene silencers, small interfering RNA/DAAO and small hairpin RNA/DAAO, almost completely prevented morphine tolerance. Intrathecal 5-chloro-benzo[d]isoxazol-3-ol and small interfering RNA/DAAO completely prevented increased spinal hydrogen peroxide levels after chronic morphine treatment. Intrathecal nonselective hydrogen peroxide scavenger phenyl-tert-N-butyl nitrone and the specific hydrogen peroxide catalyst catalase also abolished established morphine tolerance. Spinal dorsal horn astrocytes specifically expressed DAAO was significantly up-regulated, accompanying astrocyte hypertrophy after chronic morphine treatment. Conclusions: For the first time, the authors’ result identify a novel spinal astroglial DAAO–hydrogen peroxide pathway that is critically involved in the initiation and maintenance of morphine antinociceptive tolerance, and suggest that this pathway is of potential utility for the management of morphine tolerance and chronic pain.

Why is d-serine nephrotoxic and α-aminoisobutyric acid protective?

2007 ◽  
Vol 293 (1) ◽  
pp. F382-F390 ◽  
Author(s):  
Alexander W. Krug ◽  
Katharina Völker ◽  
William H. Dantzler ◽  
Stefan Silbernagl
Keyword(s):  
Reactive Oxygen Species ◽  
Amino Acid ◽  
Reactive Oxygen ◽  
Proximal Tubules ◽  
Tubular Damage ◽  
Acid Oxidase ◽  
Oxygen Species ◽  
Amino Acid Oxidase ◽  
Aminoisobutyric Acid ◽  
Serine Metabolism

d-Serine selectively causes necrosis of S3 segments of proximal tubules in rats. This leads to aminoaciduria and glucosuria. Coinjection of the nonmetabolizable amino acid α-aminoisobutyric acid (AIB) prevents the tubulopathy. d-serine is selectively reabsorbed in S3, thereby gaining access to peroxisomal d-amino acid oxidase (d-AAO). d-AAO-mediated metabolism produces reactive oxygen species. We determined the fractional excretion of amino acids and glucose in rats after intraperitoneal injection of d-serine alone or together with reduced glutathione (GSH) or AIB. Both compounds prevented the hyperaminoaciduria. We measured GSH concentrations in renal tissue before (control) and after d-serine injection and found that GSH levels decreased to ∼30% of control. This decrease was prevented when equimolar GSH was coinjected with d-serine. To find out why AIB protected the tubule from d-serine toxicity, we microinfused d-[14C]serine or [14C]AIB (0.36 mmol/l) together with [3H]inulin in late proximal tubules in vivo and measured the radioactivity in the final urine. Fractional reabsorption of d-[14C]serine and [14C]AIB amounted to 55 and 70%, respectively, and 80 mmol/l of AIB or d-serine mutually prevented reabsorption to a great extent. d-AAO activity measured in vitro (using d-serine as substrate) was not influenced by a 10-fold higher AIB concentration. We conclude from these results that 1) d-AAO-mediated d-serine metabolism lowers renal GSH concentrations and thereby provokes tubular damage because reduction of reactive oxygen species by GSH is diminished and 2) AIB prevents d-serine-induced tubulopathy by inhibition of d-serine uptake in S3 segments rather than by interfering with intracellular d-AAO-mediated d-serine metabolism.

Photo-Reactive Oxygen Species Boosting Strategy by Employing Mitochondrial Targeting Zinc-Doped Magnetic Nanoparticles to Enhance Anti-Cancer Therapy

Nano LIFE ◽  
2019 ◽  
Vol 09 (01n02) ◽  
pp. 1940005
Author(s):  
Congyu Wu ◽  
Ying Li ◽  
Jingjing Wang ◽  
Mengwei Chen ◽  
Yajing Shen ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Magnetic Nanoparticles ◽  
Cancer Therapy ◽  
Visible Region ◽  
Reactive Oxygen ◽  
Ros Generation ◽  
Oxygen Species ◽  
Mitochondrial Targeting ◽  
Intracellular Ros ◽  
Anti Cancer

The peroxidase-like functionality of iron oxide (IO) nanoparticles has attracted substantial attention in cancer treatment by reactive oxygen species (ROS) catalytic over-generation. However, the inefficient intracellular ROS generation still hurdles ready-to-use application of IO nanoparticles, attributing to the transient lifetime and limited diffusion distance of ROS. Indeed, excessive ROS generation in mitochondria is desirable to enhance cell death against cancer cells. In this study, we designed zinc-doped magnetic nanoparticles (MNPs) conjugated with triphenylphosphonium (TPP) for mitochondrial targeting. Moreover, the nanoparticles with high absorbance in visible region can catalyze ROS overproduction under visible light irradiation. Our platform provides a novel application of MNPs in targeted cancer therapy, which serves as a light-controlled switch to accelerate ROS generation and induce incremental cellular death.

Stabilization of d-amino acid oxidase from Rhodosporidium toruloides by immobilization onto magnetic nanoparticles

2008 ◽  
Vol 31 (4) ◽  
pp. 557-563 ◽  
Author(s):  
Hao-Chieh Hsieh ◽  
I-Ching Kuan ◽  
Shiow-Ling Lee ◽  
Gee-Yeng Tien ◽  
Yi-Jen Wang ◽  
...  
Keyword(s):  
Amino Acid ◽  
Magnetic Nanoparticles ◽  
Rhodosporidium Toruloides ◽  
Acid Oxidase ◽  
Amino Acid Oxidase

Purification and Characterization of Lupus Anticoagulant Like Protein from Agkistrodon Halys Brevicaudus Venom

1996 ◽  
Vol 76 (06) ◽  
pp. 0993-0997
Author(s):  
Zhao-Yan Li ◽  
Xiao-Wei Wu ◽  
Tie-Fu Yu ◽  
Eric C-Y Lian
Keyword(s):  
Amino Acid ◽  
Lupus Anticoagulant ◽  
Inhibitory Effect ◽  
Clotting Factor ◽  
Acid Oxidase ◽  
Crude Venom ◽  
Amino Acid Oxidase ◽  
Sds Page ◽  
The Individual ◽  
Reducing Condition

SummaryBy means of CM-Sephadex C-25, DEAE-Sephadex A-50, Sephadex G-200, and Sephadex G-75 chromatographies, a lupus anticoagulant like protein (LALP) from Agkistrodon halys brevicaudus was purified. On SDS-PAGE, the purified LALP had a molecular weight of 25,500 daltons under non-reducing condition and 15,000 daltons under reducing condition. The isoelectric point was pH 5.6. Its N terminal amino acid sequencing revealed a mixture of 2 sequences: DCP(P/S)(D/G)WSSYEGH(C/R)Q(Q/K). It was devoid of phospho-lipaseA, fibrino(geno)lytic, 5′-nucleotidase, L-amino acid oxidase, phosphomonoesterase, phosphodiesterase and thrombin-like activities, which were found in crude venom. In the presence of LALP, PT, aPTT, and dRVVT of human plasma were markedly prolonged and its effects were concentration-dependent but time-independent. The inhibitory effect of LALP on the plasma clotting time was enhanced by decreasing phospholipid concentration in TTI test. The individual clotting factor activity was not affected by LALP when higher dilutions of LALP-plasma mixture were used for assay. Russell’s viper venom time was shortened when high phospholipid confirmatory reagent was used. Therefore, the protein has lupus anticoagulant property.

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