The impact of heavy metals on the activity of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) kidney
Küçük Resim Yok
Tarih
2012
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
Many environmental and health problems have become a consequence of contamination of soil and water by toxic heavy metals and organic pollutants in the present age of technology. Heavy metals play vital roles in enzyme activities and other metabolic events with their bioaccumulative and nonbiodegradable properties among aquatic pollutants. Metal toxicity causes irregular metallothioneins protein synthesis, renal damage, and disruption of bone structure in humans and wildlife. In this study, we investigated in vitro effects of some metals on chemical-targeted carbonic anhydrase (CA) enzyme from rainbow trout kidney. The enzyme was purified with a specific activity of 17,285 EU × mg-1 and 31.7% yield and approximately 1800-fold using simple affinity purification method. Molecular weights of the subunit and native enzyme were estimated as 28.7 kDa and 26.9 kDa via sodium dodecyl sulfate polyacrylamide gel electrophoresis and Sephadex-G 200 column, respectively. Other kinetic properties of the enzyme were determined. Apparent Km, Vmax and kcat values were 0.40 mM, 0.097 ?mol min-1 and 15.2 s-1 for p-nitrophenylacetate substrate, respectively. Inhibitory effects of cobalt, zinc, copper, cadmium and silver on CA activity were determined using the esterase method under in vitro conditions. IC50 and Ki values were calculated for metals. Ki values for Co2+, Zn2+, Cu2+, Cd2+ and Ag+ were 0.035, 1.2, 34.8, 103 and 257 from Lineweaver-Burk graphs, respectively. Consequently, in vitro inhibition rank order was determined as Co2+ > Zn2+ > Cu 2+ > Cd2+ > Ag+. The potential inhibitor for CA was found as Co2+ from these results. © 2011 The Author(s).
Açıklama
Anahtar Kelimeler
Carbonic anhydrase, inhibition, kidney, metal toxicity, rainbow trout, 4 nitrophenylacetic acid, cadmium, carbonate dehydratase, cobalt, copper, esterase, heavy metal, metallothionein, phenylacetic acid, sephadex, silver, unclassified drug, zinc, article, binding affinity, bone structure, controlled study, enzyme activity, enzyme assay, enzyme kinetics, enzyme purification, IC 50, kidney, kidney injury, metal toxicity, molecular weight, nonhuman, polyacrylamide gel electrophoresis, protein purification, protein synthesis, rainbow trout, Animals, Carbonic Anhydrase Inhibitors, Carbonic Anhydrases, Hydrogen-Ion Concentration, Inhibitory Concentration 50, Kidney, Kinetics, Linear Models, Metals, Heavy, Oncorhynchus mykiss, Oncorhynchus mykiss, Carbonic anhydrase, inhibition, kidney, metal toxicity, rainbow trout, 4 nitrophenylacetic acid, cadmium, carbonate dehydratase, cobalt, copper, esterase, heavy metal, metallothionein, phenylacetic acid, sephadex, silver, unclassified drug, zinc, article, binding affinity, bone structure, controlled study, enzyme activity, enzyme assay, enzyme kinetics, enzyme purification, IC 50, kidney, kidney injury, metal toxicity, molecular weight, nonhuman, polyacrylamide gel electrophoresis, protein purification, protein synthesis, rainbow trout, Animals, Carbonic Anhydrase Inhibitors, Carbonic Anhydrases, Hydrogen-Ion Concentration, Inhibitory Concentration 50, Kidney, Kinetics, Linear Models, Metals, Heavy, Oncorhynchus mykiss, Oncorhynchus mykiss
Kaynak
Toxicology and Industrial Health
WoS Q Değeri
Q3
Scopus Q Değeri
Q3
Cilt
28
Sayı
4
