• Bulletin of the Korean Chemical Society
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• v.26 no.7
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• pp.1051-1056
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• 2005

The interaction of bovine carbonic anhydrase II with copper ions was studied by isothermal titration microcalorimetry, circular dichroism, UV spectrophotometry and temperature scanning spectrophotometry methods at 27 ${^{\circ}C}$ in Tris buffer solution at pH = 7.5. It was indicated that there are three non-identical different binding sites on carbonic anhydrase for $Cu^{2+}$. The binding of copper ions is exothermic and can induce some minor changes in the secondary and tertiary structure of the enzyme, which does not unfold it, but can result in a decrease in both activity and stability of the enzyme.

• The Korean Journal of Malacology
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• v.32 no.1
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• pp.63-65
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• 2016

Carbonic anhydrase (CA), which is involved in shell formation processes in bivalves, is one of the major biocatalysts for carbon capture and storage. In this study we investigated CA activity in the total hemocytic proteins of five bivalves. The highest CA activity was observed in Scapharca broughtonii, which had more than twice the activity found in Crassostrea gigas. No CA activity was observed among the total hemocytic proteins of Pinctada fucata and Saxidomus purpuratus. The results suggest that marine invertebrates may provide a better source of CA, as an alternative to mammalian sources.

• The korean journal of orthodontics
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• v.24 no.1 s.44
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• pp.115-123
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• 1994

To study bone resorption mechanism, effect of LPS on the $^{45}Ca$ release from fetal rat ulnae and radii, and effects of carbonic anhydrase inhibitors on the LPS-induced bone resorption in organ culture were studied. Ulnae and radii were removed from 19 day old fetal rats, prelabelled by subcutaneous injection of $200{\mu}Ci\;^{45}CaCl_2$ into their mother on the 17th day of gestation. Radioactivities of $^{45}Ca$ released into media were determined after 24, 48 and 72 hours. Effects of LPS and carbonic anhydrase inhibitors were observed by the ratio of $\%$ release of $^{45}Ca$ between paired control and experimental group. The observed results were as follows : 1. $LPS(1{\mu}g/ml)$ supplemented in media for 72hours increased the $^{45}Ca$ release significantly after 48 and 72 hours of culture and $LPS(10{\mu}g/m1)$ increased the $^{45}Ca$ release significantly after 72 hours of culture. 2. LPS-induced $^{45}Ca$ release was not inhibited significantly by 1mM sulfanilamide but inhibited significantly by 10mM sulfanilamide after 48 and 72 hours of culture. 3. LPS-induced $^{45}Ca$ release was not inhibited significantly by 0.1mM dichlorphenamide but inhibited significantly by 1mM dichlorphenamide after 48 and 72 hours of culture. 4. LPS-induced $^{45}Ca$ release was not inhibited significantly by 1mM acetazolamide but inhibited sighificantly by 5mM acetazolamide after 72 hours of culture.

• Journal of Life Science
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• v.24 no.2
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• pp.186-190
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• 2014

Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

• Journal of Microbiology and Biotechnology
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• v.30 no.4
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• pp.552-560
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• 2020

Human carbonic anhydrase (CA) isozyme II has been used as protein target for disorder treatment including glaucoma. Current clinically used sulfonamide-based CA inhibitors can induce side effects, and so alternatives are required. This study aimed to investigate a natural CA inhibitor from Murraya paniculata. The previously developed yeast-based assay was used to screen 14 compounds isolated from M. paniculata and identified by NMR analysis for anti-human CA isozyme II (hCAII) activity. Cytotoxicity of the compounds was also tested using the same yeast-based assay but in a different cultivation condition. Two flavonoid candidate compounds, 5, 6, 7, 8, 3', 4', 5'-heptamethoxyflavone (4) and 3, 5, 7, 8, 3', 4', 5'-heptamethoxyflavone (9), showed potent inhibitory activity against hCAII with a minimal effective concentration of 10.8 and 21.5 μM, respectively, while they both exhibited no cytotoxic effect, even at the highest concentration tested (170 μM). The results from an in vitro esterase assay of the two candidates confirmed their hCAII inhibitory activity with IC₅₀ values of 24.0 and 34.3 μM, respectively. To investigate the potential inhibition mechanism of compound 4, in silico molecular docking was performed using the FlexX and SwissDock software. This revealed that compound 4 coordinated with the Zn²⁺ ion in the hCAII active site through its methoxy oxygen at a distance of 1.60 Å (FlexX) or 2.29 Å (SwissDock). The interaction energy of compound 4 with hCAII was -13.36 kcal/mol. Thus, compound 4 is a potent novel flavonoid-based hCAII inhibitor and may be useful for further anti-CAII design and development.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.3
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• pp.527-532
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• 1992

The present paper describes temporal changes of immunohistochemical localization and quantities of carbonic anhydrase isozyme II (CA-II) prochymosin (PC) and pepsinogen (PN) in goat's abomasal mucosae during long term milk feeding. The CA-II was not detected by day 14 after birth and then became positive on day 34 in the parietal cells, suggesting that the excretion of the hydrochloric acid (HCl) begins between days 14 and 34 under a feeding condition without solid materials. The quantity of the PC in the gastric chief cells detected by the ELISA showed rapid increase from the day of birth, making a peak on day 8 and then gradually decreased with age. The decrease in quantity of PC became started during the time period when HCl excretion had not started yet. The quantities of PN in the gastric chief cells were almost stable during the whole period examined. Expressions of these gastric enzymes did not seem to be regulated by the change of feeding condition.

• Particle and aerosol research
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• v.9 no.4
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• pp.201-208
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• 2013

Copper (II) and Nickel (II) mimic complexes of enzyme carbonic anhydrase were evaluated under ambient condition for carbon dioxide capture and conversion process. The synthesized complexes were characterized by ATR-FTIR and UV-DR spectroscopy. It was found that all the complexes have biomimetic activity towards $CO_2$ using para-nitrophenyl acetate (p-NPA) hydrolysis as the model reaction. Interestingly, the proper geometry obtained by the restricted orientation of tripodal N atoms in Cu (II) complex of 2,6-bis(2-benzimidazolyl) pyridine showed the highest activity (1.14 au) compared to others. The $CO_2$ bio-mineralization to $CaCO_3$ was carried out via in-vitro crystallization approach. Results indicate that the biomimetic complexes have a role in determining $CaCO_3$ morphology. The present observations establish a qualitative insight for the design of improved small-molecule catalysts for carbon capture.

• BMB Reports
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• v.40 no.2
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• pp.205-211
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• 2007

The stability curve - a plot of the Gibbs free energy of unfolding versus temperature - is calculated for bovine erythrocyte carbonic anhydrase in 150 mM sodium phosphate (pH = 7.0) from a combination of reversible differential scanning calorimetry measurements and isothermal guanidine hydrochloride titrations. The enzyme possesses two stable folded conformers with the conformational transition occurring at ~30$^{\circ}C$. The methodology yields a stability curve for the complete unfolding of the enzyme below this temperature but only the partial unfolding, to the molten globule state, above it. The transition state thermodynamics for the low- to physiological-temperature conformational change are calculated from slow-scan-rate differential scanning calorimetry measurements where it is found that the free energy barrier for the conversion is 90 kJ/mole and the transition state possesses a substantial unfolding quality. The data therefore suggest that the x-ray structure may differ considerably from the physiological structure and that the two conformers are not readily interconverted.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.1
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• pp.351-354
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• 2013

Objective: To assess the practical utility of pleural fluid carbonic anhydrase XII (CAXII) quantification for differential diagnosis of effusions. Materials and Methods: Fluid was collected prospectively from fifty patients presenting with lymphocytic pleural effusions for investigation and CAXII was quantified by ELISA. Results: Pleural fluid CAXII concentrations were significantly higher in lung cancer patients (n=30) than in tuberculous controls (n=20). The sensitivity and specificity of this biomarker were 60%and 75%, respectively. CAXII measurement was not inferior to cytological examination in the diagnosis and exclusion of pleural effusions from lung cancer patitents (sensitivity 60% vs. 57%; specificity 75% vs. 100%; positive predictive value 77%; negative predictive value 54%). In patients with negative cytology, it offered a sensitivity of 54%. Conclusions: Pleural fluid CAXII is elevated in pleural effusions from lung cancer patients. Measurement of CAXII may be used in the future as a valuable adjunct to cytology in the diagnostic assessment of patients with pleural effusions related to lung cancer, especially when cytological examination is inconclusive.

• BMB Reports
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• v.39 no.5
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• pp.636-641
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• 2006

Our previous studies indicated that native carbonic anhydrase does not interact with hydrophobic adsorbents and that it acquires this ability upon denaturation. In the present study, an apo form of the enzyme was prepared by removal of zinc and a comparative study was performed on some characteristic features of the apo and native forms by far- and near-UV circular dichroism (CD), intrinsic fluorescent spectroscopy, 1-anilino naphthalene-8-sulfonate (ANS) binding, fluorescence quenching by acrylamide, and Tm measurement. Results indicate that protein flexibility is enhanced and the hydrophobic sites become more exposed upon conversion to the apo form. Accordingly, the apo structure showed a greater affinity for interaction with hydrophobic adsorbents as compared with the native structure. As observed for the native enzyme, heat denaturation of the apo form promoted interaction with alkyl residues present on the adsorbents and, by cooling followed by addition of zinc, catalytically-active immobilized preparations were obtained.