Summary
The interaction of Cd2+ with the plasma membrane Ca2+-transporting ATPase of fish gills was studied. ATP-driven Ca2+-transport in basolateral membrane (BLM) vesicles was inhibited by Cd2+ with anI 50 value of 3.0nm at 0.25 μm free Ca2+ using EGTA, HEEDTA and NTA to buffer Ca2+ and Cd2+ concentrations. The inhibition was competitive in nature since theK 0.5 value for Ca2+ increased linearly with increasing Cd2+ concentrations while theV max remained unchanged. The Ca2+ pump appeared to be calmodulin dependent, but we conclude that the inhibition by Cd2+ occurs directly on the Ca2+ binding site of the Ca2+-transporting ATPase and not via the Ca2+-binding sites of calmodulin. It is suggested that Cd2+-induced inhibition of Ca2+-transporting enzymes is the primary effect in the Cd2+ toxicity towards cells followed by several secondary effects due to a disturbed cellular Ca2+ metabolism. Our data illustrate that apparent stimulatory effects of low concentrations of Cd2+ on Ca2+-dependent enzymes may derive from increased free-Ca2+ levels when Cd2+ supersedes Ca2+ on the ligands.
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Akerman, K.E.O., Honkaniemi, J., Scott, I.G., Andersson, L.C. 1985. Interaction of Cd2+ with the calmodulin-activated (Ca2++Mg2+)-ATPase activity of human erythrocyte ghosts.Biochim. Biophys. Acta 845:48–53
Benoit, D.A., Leonhard, E.N., Christensen, G.M., Fiandt, J.T. 1976. Toxic effects of cadmium on three generations of brook trout (Salvelinus fontinalis).Trans. Am. Fish. Soc. 105:550–560
Berg, A. 1970. Studies on the metabolism of calcium and strontium in freshwater fish. II. Relative contribution of direct and intestinal absorption in growth conditions.Mem. Ist. Ital. Idrobiol. Dott Marco Marchi 26:241–255
Bishop, W.E., McIntosh, A.W. 1981. Acute lethality and effects of sublethal cadmium exposure on ventilation frequency and cough rate of bluegill (Lepomus macrochirus).Arch. Contam. Toxicol. 10:519–530
Cain, K., Webb, M. 1983. Metallothionein and its relationship to the toxicity of cadmium and other metals in the young rat. In: Health Evaluation of Heavy Metals in Infant Formula and Junior Food. E.H.F. Schmidt and A.G. Hildenbrandt, editors. pp. 105–111. Springer, Berlin, Heidelberg
Caroni, P., Carafoli, E. 1981. The Ca2+-pumping ATPase of heart sarcolemma.J. Biol. Chem. 256:3263–3270
Chao, S.H., Suzuki, Y., Zysk, J.R., Cheung, W.Y. 1984. Activation of calmodulin by various metal cations as a function of ionic radius.Mol. Pharmacol. 26:75–82
Christensen, G.M. 1975. Biochemical effects of methylmercuric chloride, cadmium chloride and lead nitrate on embryos and alevins of the brook trout,Salvelinus fontinalis.Toxicol. Appl. Pharmacol. 32:191–197
Flik, G., Van de Winkel, J.G.J., Part, P., Wendelaar Bonga, S.E., Lock, R.A.C. 1987. Calmodulin-mediated cadmium inhibition of phosphodiesterase activity, in vitro.Arch. Toxicol. 59:353–359
Flik, G., Van Rijs, J.H., Wendelaar Bonga, S.E. 1985a. Evidence for high affinity and ATP-driven Ca2+-transport in membrane preparations of the gill epithelium of the cichlid fishOreochromis mossambicus.J. Exp. Biol. 119:335–347
Flik, G., Wendelaar Bonga, S.E., Fenwick, J.C. 1983. Ca2+-dependent phosphatase and ATPase activities in eel gill plasma membranes. I. Identification of Ca2+-activated ATPase activities with non-specific phosphatase activities.Comp. Biochem. Physiol. 76B:745–754
Flik, G., Wendelaar Bonga, S.E., Fenwick, J.C. 1984. Ca2+-dependent phosphatase and Ca2+-dependent ATPase activities in plasma membranes of eel gill epithelium. II. Evidence for transport high affinity Ca2+-ATPase.Comp. Biochem. Physiol. 79B:9–16
Flik, G., Wendelaar Bonga, S.E., Fenwick, J.C. 1985b. Active Ca2+-transport in plasma membranes of branchial epithelium of the North American eelAnguilla rostrata LeSueur.Biol. Cell 55:265–272
Foder, B., Scharff, O. 1981. Decrease of apparent calmodulin affinity of erythrocyte (Ca2+/Mg2+)-ATPase at low Ca2+-concentrations.Biochim. Biophys. Acta 649:367–376
Forsen, S., Thulin, E., Drakenberg, T., Krebs, J., Seamon, K. 1980. A113Cd NMR study of calmodulin and its interaction with calcium, magnesium and trifluoperazine.FEBS Lett. 117:189–194
Ghijsen, W.E.J.M., Van Os, C.H., Heizmann, C.W., Murer, H. 1986. Regulation of duodenal Ca2+ pump by calmodulin and vitamin D-dependent Ca2+-binding protein.Am. J. Physiol. 251:G223-G229
Giles, M.A. 1984. Electrolyte and water balance in plasma and urine of rainbow trout (Salmo gairdneri) during chronic exposure to cadmium.Can. J. Fish. Aquat. Sci. 41:1678–1685
Ichii, T., Mugiya, Y. 1983. Effects of a dietary deficiency in calcium on growth and calcium uptake from the aquatic environment in the goldfish,Carassius auratus.Comp. Biochem. Physiol. 74A:259–262
Larsen, F.L., Vincenzi, F.F. 1979. Calcium transport across the plasma membrane: Stimulation by calmodulin.Science 204:306–309
Lynch, T.J., Cheung, W.Y. 1979. Human erythrocyte Ca2+-Mg2+-ATPase: Mechanism of stimulation by Ca2+.Arch. Biochem. Biophys. 194:165–170
McCarty, L.S., Houston, A.H. 1976. Effects of exposure to sublethal levels of cadmium upon water-electrolyte status in the goldfish (Carassius auratus).J. Fish Biol. 9:11–19
Maitani, T., Watahiki, A., Suzuki, K.T. 1986. Acute renal dysfunction by cadmium injected with cysteine in relation to renal critical concentration of cadmium.Arch. Toxicol. 58:136–140
Majewski, H.S., Giles, M.A. 1981. Cardiovascular-respiratory response of rainbow trout (Salmo gairdneri) during chronic exposure to sublethal concentrations of cadmium.Water Res. 15:1211–1217
Mazzei, G.J., Girard, P.R., Kuo, J.F. 1984. Environmental pollutant Cd2+ biphasically and differentially regulates myosin light chain kinase and phospholipid/Ca2+-dependent protein kinase.FEBS Lett. 173:124–128
Milet, C., Peignoux-Deville, J., Martelly, E. 1979. Gill calcium flows in the eel,Anguilla anguilla (L). Effects of Stannius corpuscles and ultimo-branchial body.Comp. Biochem. Physiol. 63A:63–70
Nechay, B.R., Saunders, J.P. 1977. Inhibition of renal adenosine triphosphate by cadmium.J. Pharmacol. Exp. Ther. 200:623–629
Payan, P., Matty, A. 1975. The characteristics of ammonia excretion by a perfused isolated-head of trout (Salmogairdneri): Effect of temperature and CO2-free Ringer.J. Comp. Physiol. 96:167–184
Pecoraro, V.L., Hermes, J.D., Cleland, W.W. 1984. Stability constants of Mg2+ and Cd2+ complexes of adenine nucleotides and thionucleotides and rate constants for formation and dissociation of MgATP and MgADP.Biochemistry 23:5262–5271
Perry, S.F., Flik, G. 1987. A characterization of branchial transepithelial calcium fluxes in the freshwater trout (Salmogairdneri).Am. J. Physiol. 254:R491-R498
Price, N.C., Dwek, R.A. 1979. Binding of ligands to macromolecules. In: Principles and Problems in Physical Chemistry for Biochemists. Second edition. pp. 32–36. Clarendon, Oxford
Rauchova, H., Kaul, P.P., Drahota, Z. 1985. Activation of mitochondrial glycerol 3-phosphate dehydrogenase by cadmium ions.Gen. Physiol. Biophys. 4:29–34
Roch, M., Maly, E.J. 1979. Relationship of cadmium-induced hypocalcemia with mortality in rainbow trout (Salmo gairdneri) and the influence of temperature on toxicity.J. Fish. Res. Board Can. 36:1297–1303
Sillen, L.G., Martell, A.E. 1964. Stability constants of metal ion complexes.The Chemical Society, Spec. Publ. no. 17, London
Sugawara, N., Sugawara, C. 1975. Effect of cadmium,in vivo andin vitro, on intestinal brush border ALPase and ATPase.Bull. Environ. Contam. Toxicol. 14:653–656
Suzuki, Y., Chao, S.H., Zysk, J.R., Cheung, W.Y. 1985. Stimulation of calmodulin by cadmium ion.Arch. Toxicol. 57:205–211
Van Heeswijk, M.P.E., Geertsen, J.A.M., Van Os, C.H. 1984. Kinetic properties of the ATP-dependent Ca2+ pump and the Na+/Ca2+ exchange system in basolateral membranes from rat kidney cortex.J. Membrane Biol. 79:19–31
Van Os, C.H. 1987. Transcellular calcium transport in intestinal and renal epithelial cells.Biochim. Biophys. Acta 906:195–222
Verbost, P.M., Flik, G., Lock, R.A.C., Wendelaar Bonga, S.E. 1987a. Cadmium inhibition of Ca2+-uptake in rainbow trout gills.Am. J. Physiol. 253:R216-R221
Verbost, P.M., Senden, M.H.M.N., Van Os, C.H. 1987b. Nanomolar concentrations of Cd2+ inhibit Ca2+ transport systems in plasma membranes and intracellular Ca2+ stores in intestinal epithelium.Biochim. Biophys. Acta 902:247–252
Webb, M. 1979. 2. The metallothioneins. In: Topics in Environmental Health. M. Webb, editor. pp. 195–266. Elsevier/North Holland Biomedical, New York
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Verbost, P.M., Flik, G., Lock, R.A.C. et al. Cadmium inhibits plasma membrane calcium transport. J. Membrain Biol. 102, 97–104 (1988). https://doi.org/10.1007/BF01870448
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DOI: https://doi.org/10.1007/BF01870448