Abstract
The effect of regucalcin, a novel Ca2+-binding protein, on Ca2+/ calmodulin-dependent cyclic adenosine monophosphate (AMP) phosphodiesterase activity in the cytosol of rat renal cortex was investigated. Regucalcin with physiologic concentration (10-7 M) in rat kidney had no effect on cyclic AMP phosphodiesterase activity in the absence of CaCl2 and calmodulin. However, the activatory effect of both CaCl2 (10 µM) and calmodulin (20 U/ml) on cyclic AMP phosphodiesterase was markedly inhibited by the addition of regucalcin (10-8 to 10-6 M) in the enzyme reaction mixture. The inhibitory effect of regucalcin on the enzyme activity was also seen in the presence of CaCl2 (5-50 µM) or calmodulin (5-50 U/ml) with increasing concentrations. The presence of trifluoperazine (10 µM), an antagonist of calmodulin, caused a partial inhibition of Ca2+ /calmodulin-dependent cyclic AMP phosphodiesterase activity. This inhibition was further enhanced by the addition of regucalcin (10-7 M). The inhibitory effect of regucalcin (10-7 M) was not seen in the presence of 20 µM trifluoperazine. Moreover, the activatory effect of calmodulin (20 U/ml) on cyclic AMP phosphodiesterase was not entirely seen, when calmodulin was added 10 min after incubation in the presence of CaCl2 (10 µM) and regucalcin (10-7 M). The present results demonstrates that regucalcin has an inhibitory effect on Ca2+ /calmodulin-dependent cyclic AMP phosphodiesterase activation in the cytosol of rat renal cortex.
Similar content being viewed by others
References
Yamaguchi M: Physicochemical properties of calcium-binding protein isolated from rat liver cytosol: Ca2+-induced conformational changes. Chem Pharm Bull 36: 286–290, 1988
Shimokawa N, Yamaguchi M: Molecular cloning and sequencing of the cDNA coding for a calcium-binding regucalcin from rat liver. FEBS Lett 327: 251–255, 1993
Shimokawa N, Matsuda Y, Yamaguchi M: Genomic cloning and chromosomal assignment of rat regucalcin gene. Mol Cell Biochem 151: 157–163, 1995
Shimokawa N, Yamaguchi M: Calcium administration stimulates the expression of calcium-binding protein regucalcin mRNA in rat liver. FEBS Lett 305: 151–154, 1992
Yamaguchi M, Kurota H: Expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats: The stimulation by calcium administration. Mol Cell Biochem 146: 71–77, 1995
Yamaguchi M: A novel Ca2+-binding protein regucalcin and calcium inhibition: Regulatory role in liver cell function. In: K. Kohama (ed). Calcium Inhibition. Japan Sci Soc Press, Tokyo and CRC Press, Boca Raton, 1992, pp 19–41
Mori S, Yamaguchi M: Hepatic calcium-binding protein regucalcin decreases Ca2+ /calmodulin-dependent protein kinase activity in rat liver cytosol. Chem Pharm Bull 38: 2216–2218, 1990
Yamaguchi M, Mori S: Inhibitory effect of calcium-binding protein regucalcin on protein kinase C activity in rat liver cytosol. Biochem Med Metab Biol 43: 140–146, 1990
Kurota H, Yamaguchi M: Activatory effect of calcium-binding protein regucalcin on ATP-dependent calcium transport in the basolateral membranes of rat kidney cortex. Mol Cell Biochem 169: 149–156, 1997
Yamaguchi M, Yamamoto T: Purification of calcium binding substance from soluble fraction of normal rat liver. Chem Pharm Bull 26: 1915–1918, 1978
Kakiuchi S, Yamazaki R, Teshima Y, Uenishi K, Miyamoto E: Multiple cyclic nucleotide phosphodiesterase activities from rat tissues and occurrence of a calcium-plus magnesium-ion-dependent phospho-diesterase and its protein activator. Biochem J 146: 109–120, 1975
Butcher RW, Sutherland EW: Adenosine 3′,5′-phosphate in biological materials. I. Purification and properties of cyclic 3′,5′-nucleotide phosphodiesterase and use of this enzyme to characterize adenosine 3′, 5′-phosphate in human urine. J Biol Chem 237: 1244–1250, 1962
Nakamura M, Mori K: Colorimetric determination of inorganic phosphorus in the presence of glucose-l-phosphate and adenosine triphosphate. Nature 182: 1441–1442, 1958
Lowry OH, Rosebrough NJ, Farr AL, Randall FJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–273, 1951
Kakiuchi S, Yasuda S, Yamazaki R, Teshima Y, Kanda K, 16 Kakiuchi R, Sobue K: Quantitative determinations of calmodulin in the supernatant and particulate fractions of mammalian tissues. J Biochem 92: 1041–1048, 1982
Rasmussen J: Cell communication, calcium ion, and cyclic adenosine monophosphate. Science 170: 404–412, 1970
Cheung WY: Calmodulin plays a pivotal role in cellular regulation. Science 202: 19–27, 1980
Williamson JR, Cooper RH, Hoek JB: Role of calcium in the hormonal regulation of liver metabolism. Biochim Biophys Acta 639: 243–295, 1981
Reinhart PH, Taylor WM, Bygrave FL: The role of calcium ion in the mechanisms of action of a-adrenergic agonists in rat liver. Biochem J 223: 1–13, 1984
Kitamura K, Miller RT: Regulation of hormone-sensitive calcium influx by the adenylyl cyclase system in renal epithelial cells. J Clin Invest 94: 328–336, 1994
Azarani A, Goltzman D, Orlowski J: Parathyroid hormone and parathyroid hormone-related peptide inhibit the apical Na+ /H+ exchanger NHE-3 isoform in renal cells (OK) via a dual signaling cascade involving protein kinase A and C. J Biol Chem 270: 20004–20010, 1995
Yamaguchi M, Isogai M: Tissue concentration of calcium-binding protein regucalcin in rats by enzyme-linked immunoadsorbent assay. Mol Cell Biochem 122: 65–68, 1993
Yamaguchi M, Sugii K: Properties of calcium-binding protein isolated from the soluble fraction of normal rat liver. Chem Pharm Bull 29: 567–570, 1981
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Yamaguchi, M., Kurota, H. Inhibitory effect of regucalcin on Ca2+/calmodulin-dependent cyclic AMP phosphodiesterase activity in rat kidney cytosol. Mol Cell Biochem 177, 209–214 (1997). https://doi.org/10.1023/A:1006829926590
Issue Date:
DOI: https://doi.org/10.1023/A:1006829926590