Characterization of a high activity carbonic anhydrase isozyme purified from erythrocytes of Salmo gairdneri

Hall, Greg; Schraer, Rosemary

doi:10.1016/0305-0491(83)90043-3

Cited by 14 publications

(13 citation statements)

References 30 publications

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“…This result of MW was similar Fig. 4 K i graphs for carbonic anhydrase from rainbow trout brain a, b, c, d and e Lineweaver-Burk graphs in five different substrate (p-nitrophenylacetate) concentrations and in three different Co 2+ , Cu 2+ , Zn 2+ , Ag + , and Cd 2+ concentrations for determination of K i for Co 2+ , Cu 2+ , Zn 2+ , Ag + , and Cd 2+ , respectively to CAs purified from many other tissues such as Salmo gairdneri erythrocyte, flounder gills, Pisum sativum [31,33,34]. However, as can be seen from the literature, the molecular weight of the carbonic anhydrase has showed a variety among different tissues [35,36].…”

Section: Discussionmentioning

confidence: 99%

“…In literature, the enzyme has been purified by different methods. For example, the enzyme was purified 3.7-fold from 300 ml blood sample using the chloroform-ethanol extraction, Sephadex G-75 gel filtration column and DEAE Bio-Gel ion exchange material from RT erythrocytes [31]. Krungkrai et al [15] purified the enzyme from Plasmodium falciparum with 66.7-fold purification and 18 EU/mg protein using combinations of cation-exchange mono S column, anion-exchange mono Q column, and Sepharose 12 gel filtration column.…”

Section: Discussionmentioning

confidence: 99%

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Effects of Some Metals on Carbonic Anhydrase from Brains of Rainbow Trout

Söyüt

Beydemır

Hisar

2008

Biol Trace Elem Res

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Carbonic anhydrase (CA) enzyme was purified from rainbow trout brain by Sepharose-4B-L: -tyrosine-sulfanilamide affinity chromatography. The enzyme was obtained with a specific activity of 2,275 EU mg(-1) and a yield of 22.5%. The sample obtained from the affinity column was used for kinetic properties and inhibition studies. Both optimum and stable pH were found as 9.0 in 1 M Tris-SO(4) at 4 degrees C, respectively. To check the purity and subunit molecular weight of enzyme, sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis was performed, and MW was found as approximately 29.0 kDa. The molecular weight of native enzyme was estimated to be approximately 27.3 kDa by gel filtration chromatography. The purified enzyme had apparent K (m),V (max), and k (cat) as follows: 0.92 mM, 0.207 micromol.min(-1) and 43.6 s(-1) for p-nitrophenylacetate. The inhibitory effects of Co(II), Cu(II), Zn(II), Ag(I), and Cd(II) on CA enzyme activity were determined using the esterase method under in vitro conditions at low concentrations of the corresponding metals. The obtained IC(50) values, which cause 50% inhibition on in vitro enzyme activity, were 0.05, 30, 0.31, 159, and 82.5 mM for cobalt, copper, zinc, silver, and cadmium, respectively. K ( i ) values were also calculated from Linewaever-Burk plots for these substances as 0.014, 27.68, 2.15, 193.86, and 94.18 for cobalt, copper, zinc, silver, and cadmium, respectively; it was determined that cobalt, silver and cadmium inhibited the enzyme competitively, copper inhibited noncompetitively while zinc inhibited the enzyme uncompetitively.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Effects of Some Metals on Carbonic Anhydrase from Brains of Rainbow Trout

Söyüt

Beydemır

Hisar

2008

Biol Trace Elem Res

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“…Different chromatography methods have been used to purify CA in fish (Hall and Schraer, 1983). However, these methods such as Sephadex G-75 gel filtration and DEAE Bio Gel anion exchange chromatography are very laborious and time-consuming.…”

Section: Discussionmentioning

confidence: 99%

Effect of vitamin E on carbonic anhydrase enzyme activity in rainbow trout (Oncorhynchus mykiss) erythrocytes in vitro and in vivo

Aras-Hisar

Hisar

Beydemır

et al. 2004

Acta Veterinaria Hungarica

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Considering that the excessive usage of vitamin E causes hypervitaminosis and thus reduces blood erythrocyte concentrations, therefore it is worth studying how its pharmacological dosage affects the activity of carbonic anhydrase (CA) enzyme found in erythrocytes of rainbow trout (Oncorhynchus mykiss) in vitro and in vivo. Vitamin E inhibited CA enzyme and the IC 50 value of the vitamin was 0.039 mM in vitro. Similarly, it was seen that vitamin E inhibited CA enzyme activity after the first hour following vitamin E injections in vivo. The activities of CA in groups of trout given vitamin E injection were measured at 1, 3 and 5 h and the corresponding activities were found to be 772.7 ± 290.5 (P < 0.05), 1286.4 ± 378.2 and 1005.7 ± 436.1 enzyme units (EU) g Hb -1 . The difference over the control was significant (P < 0.05) in the first hour and insignificant at 3 and 5 h (P > 0.05). The activity of CA in the control, which did not contain vitamin E, was determined as 1597.7 ± 429.0 EU g Hb -1 .

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“…Salmo gairdneri CA was purified by Hall and Schraer using three purification steps: chloroform ethanol extraction, size exclusion chromatography (Sephadex G-75) and anion exchange chromatography (DEAE Bio-Gel) 52 . As described in the literature, for a large-scale preparation of bovine and porcine CAs was used the rapid ion-exchange chromatography technique on DEAEcellulose column to obtain the catalysts in its pure form.…”

Section: Smentioning

confidence: 99%

A magnificent enzyme superfamily: carbonic anhydrases, their purification and characterization

Guler

Capasso

Supuran

2015

Journal of Enzyme Inhibition and Medicinal Chemistry

130

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In this paper, we reviewed the purification and characterization methods of the a-carbonic anhydrase (CA, EC 4.2.1.1) class. Six genetic families (a-, b-, g-, d-, z-and Z-CAs) all know to date, all encoding such enzymes in organisms widely distributed over the phylogenetic tree. Starting from the manuscripts published in the 1930s on the isolation and purification of a-CAs from blood and other tissues, and ending with the recent discovery of the last genetic family in protozoa, the Z-CAs, considered for long time an a-CA, we present historically the numerous and different procedures which were employed for obtaining these catalysts in pure form. a-CAs possess important application in medicine (as many human a-CA isoforms are drug targets) as well as biotechnological processes, in which the enzymes are ultimately used for CO 2 capture in order to mitigate the global warming effects due to greenhouse gases. Recently, it was discovered an involvement of CAs in cancerogenesis as well as infection caused by pathogenic agents such as bacteria, fungi and protozoa. Inhibition studies of CAs identified in the genome of the aforementioned organisms might lead to the discovery of innovative drugs with a novel mechanism of action.

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Characterization of a high activity carbonic anhydrase isozyme purified from erythrocytes of Salmo gairdneri

Cited by 14 publications

References 30 publications

Effects of Some Metals on Carbonic Anhydrase from Brains of Rainbow Trout

Effects of Some Metals on Carbonic Anhydrase from Brains of Rainbow Trout

Effect of vitamin E on carbonic anhydrase enzyme activity in rainbow trout (Oncorhynchus mykiss) erythrocytes in vitro and in vivo

A magnificent enzyme superfamily: carbonic anhydrases, their purification and characterization

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