Isolation, purification and characterization of an insect carbonic anhydrase

Burt, E.; Darlington, Mark V.; Graf, George; Meyer, Haruko

doi:10.1016/0965-1748(92)90066-n

Cited by 10 publications

(4 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition to this, CA has been purified and its kinetic properties have been examined from a wide range of sources including human erythrocytes [11], fish gills [12], fish erythrocytes [13], rat saliva, rat erythrocytes [14], Plasmodium falciparum [15], and insects [16]. Besides, it has been reported that CA was partially characterized from plant, yeast, and bacteria [15].…”

mentioning

confidence: 99%

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

Söyüt

Beydemır

Hisar

2008

Biol Trace Elem Res

View full text Add to dashboard Cite

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.

show abstract

mentioning

confidence: 99%

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

Söyüt

Beydemır

Hisar

2008

Biol Trace Elem Res

View full text Add to dashboard Cite

show abstract

“…The enzyme has been purified from many tissues including human erythrocytes and its kinetic properties have been examined in human erythrocytes [6], fish gills [7] and erythrocytes [8], rat saliva and erythrocytes [9], Plasmodium falciparum [10], insect [11], bovine bone [12], bovine leucocytes [13]. Morover, it has been reported that CA has been partially characterized from plants, yeast and bacteria [10].…”

Section: Introductionmentioning

confidence: 99%

In vitroinhibitory effects of some heavy metals on human erythrocyte carbonic anhydrases

Ekinci

Beydemır

Küfrevioğlu

2007

Journal of Enzyme Inhibition and Medicinal Chemistry

View full text Add to dashboard Cite

The inhibition of two human carbonic anhydrase (HCA, EC 4.2.1.1) isozymes, the cytosolic HCA I and II, with heavy metal salts of Pb(II), Co(II) and Hg(II)has been investigated. Human erythrocyte CA-I isozyme was purified with a specific activity of 920 EUmg 21 and a yield of 30% and CA-II isozyme was purified with a specific activity of 8000 EUmg 21 and a yield of 40% using Sepharose-4B-L tyrosine-sulfanilamide affinity gel chromatography. The overall purification was approximately 104-fold for HCA-I and 900-fold for HCA-II. The inhibitory effects of different heavy metals (lead, cobalt and mercury) on CA activity were determined at low concentrations using the esterase method under in vitro conditions. K i values for these metals were calculated from Lineweaver-Burk graphs as 1.0, 3.22 and 1.45 mM for HCA-I and 0.059, 1.382 and 0.32 mM for HCA-II respectively. Lead was a noncompetitive inhibitor for HCA-I and competitive for HCA-II, cobalt was competitive for HCA-I and noncompetitive for HCA-II and mercury was uncompetitive for both HCA-I and HCA-II. Lead was the best inhibitor for both HCA-I and HCA-II.

show abstract

“…Carbonic anhydrases are metabolic enzymes whose main role is to catalyse the reversible hydratation of CO 2 ( 76 , 77 ). They are responsible for maintaining the correct pH in insect tissues.…”

Section: Discussionmentioning

confidence: 99%

Immune priming modulates Galleria mellonella and Pseudomonas entomophila interaction. Antimicrobial properties of Kazal peptide Pr13a

Sułek,

Kordaczuk,

Mak

et al. 2024

Front. Immunol.

View full text Add to dashboard Cite

Galleria mellonella larvae repeatedly infected with Pseudomonas entomophila bacteria re-induced their immune response. Its parameters, i.e. the defence activities of cell-free hemolymph, the presence and activity of antimicrobial peptides, and the expression of immune-relevant genes were modulated after the re-challenge in comparison to non-primed infected larvae, resulting in better protection. No enhanced resistance was observed when the larvae were initially infected with other microorganisms, and larvae pre-infected with P. entomophila were not more resistant to further infection with other pathogens. Then, the peptide profiles of hemolymph from primed- and non-primed larvae infected with P. entomophila were compared by quantitative RP-HPLC (Reverse Phase - High Performance Liquid Chromatography). The level of carbonic anhydrase, anionic peptide-1, proline peptide-2, and finally, unknown so far, putative Kazal peptide Pr13a was higher in the primed infected animals than in the larvae infected with P. entomophila for the first time. The expression of the Pr13a gene increased two-fold after the infection, but only in the primed animals. To check whether the enhanced level of Pr13a could have physiological significance, the peptide was purified to homogeneity and checked for its defence properties. In fact, it had antibacterial activity: at the concentration of 15 µM and 7.5 µM it reduced the number of P. entomophila and Bacillus thuringiensis CFU, respectively, to about 40%. The antibacterial activity of Pr13a was correlated with changes observed on the surface of the peptide-treated bacteria, e.g. surface roughness and adhesion force. The presented results bring us closer to finding hemolymph constituents responsible for the effect of priming on the immune response in re-infected insects.

show abstract

Isolation, purification and characterization of an insect carbonic anhydrase

Cited by 10 publications

References 23 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

In vitroinhibitory effects of some heavy metals on human erythrocyte carbonic anhydrases

Immune priming modulates Galleria mellonella and Pseudomonas entomophila interaction. Antimicrobial properties of Kazal peptide Pr13a

Contact Info

Product

Resources

About