Characterization of putrescine and cadaverine export in mammalian cells. A pharmacological approach.

We characterized the mechanism(s) involved in the efflux of putrescine/cadaverine from cultured mammalian cells using various pharmacological agents. Verapamil and quinine inhibited putrescine and cadaverine export in monocytic-leukemic RAW 264 and H35 hepatoma cells in a concentration-dependent manner with an IC50 in the micromolar range. Verapamil, which inhibits L-type calcium channels, inhibited putrescine export, regardless of whether calcium was present in the extracellular medium or not. Furthermore, the export of putrescine in the absence of verapamil did not appear to depend upon extracellular calcium. Neither intracellular calcium, external sodium, changes in intracellular pH nor phosphorylation affected the levels of putrescine export independently from changes in intracellular putrescine levels. The data suggest that verapamil and quinine inhibit putrescine/cadaverine efflux from the cell by binding directly to an integral membrane protein.

[1]  W. Gahl,et al.  Polyamine degradation in foetal and adult bovine serum. , 1982, The Biochemical journal.

[2]  A. Borle,et al.  Effects of pH on Ca2+i, Na+i, and pHi of MDCK cells: Na(+)-Ca2+ and Na(+)-H+ antiporter interactions. , 1991, The American journal of physiology.

[3]  L. Marton,et al.  Solid-phase extraction and determination of dansyl derivatives of unconjugated and acetylated polyamines by reversed-phase liquid chromatography: improved separation systems for polyamines in cerebrospinal fluid, urine and tissue. , 1986, Journal of chromatography.

[4]  R. Smallridge,et al.  Na+-H+ antiport and monensin effects on cytosolic pH and iodide transport in FRTL-5 rat thyroid cells. , 1992, The American journal of physiology.

[5]  I. Pastan,et al.  Certain calcium channel blockers bind specifically to multidrug-resistant human KB carcinoma membrane vesicles and inhibit drug binding to P-glycoprotein. , 1987, The Journal of biological chemistry.

[6]  E. R. Seidel,et al.  Polyamine-dependent growth and calmodulin-regulated induction of ornithine decarboxylase. , 1989, The American journal of physiology.

[7]  J. Daugirdas,et al.  Cytosolic free calcium regulation in response to acute changes in intracellular pH in vascular smooth muscle. , 1993, The American journal of physiology.

[8]  K. Longenecker,et al.  Mechanism of putrescine transport in human pulmonary artery endothelial cells. , 1993, Journal of Pharmacology and Experimental Therapeutics.

[9]  J. Ristow,et al.  Uptake, intracellular binding, and excretion of polyamines during growth of Neurospora crassa. , 1989, Archives of biochemistry and biophysics.

[10]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[11]  T. Tsuruo,et al.  Phosphorylation of the Mr 170,000 to 180,000 glycoprotein specific to multidrug-resistant tumor cells: effects of verapamil, trifluoperazine, and phorbol esters. , 1987, Cancer research.

[12]  M. Volm,et al.  Activity of various amphiphilic agents in reversing multidrug resistance of L 1210 cells. , 1990, Cancer letters.

[13]  A. Yoshida,et al.  Role of calcium ion in induction of apoptosis by etoposide in human leukemia HL-60 cells. , 1993, Biochemical and biophysical research communications.

[14]  Rowland H. Davis.  Management of polyamine pools and the regulation of ornithine decarboxylase , 1990, Journal of cellular biochemistry.

[15]  Y. Nishizuka,et al.  Inhibitory action of chlorpromazine, dibucaine, and other phospholipid-interacting drugs on calcium-activated, phospholipid-dependent protein kinase. , 1980, The Journal of biological chemistry.

[16]  S. Otani,et al.  Two mechanisms of spermidine/spermine N1-acetyltransferase-induction. , 1989, Archives of biochemistry and biophysics.

[17]  A. Pegg,et al.  Polyamine metabolism and its importance in neoplastic growth and a target for chemotherapy. , 1988, Cancer research.

[18]  W. Dalton,et al.  Synergistic inhibition by verapamil and quinine of P-glycoprotein-mediated multidrug resistance in a human myeloma cell line model. , 1991, Blood.

[19]  J. Endicott,et al.  The biochemistry of P-glycoprotein-mediated multidrug resistance. , 1989, Annual review of biochemistry.

[20]  M. Grillo,et al.  Efflux of polyamines from human lymphocytes and from L 1210 cells. , 1992, The International journal of biochemistry.

[21]  H L Pearce,et al.  Physical-chemical properties shared by compounds that modulate multidrug resistance in human leukemic cells. , 1988, Molecular pharmacology.

[22]  S. Yamamoto,et al.  Protein kinase C-dependent and -independent actions of a potent protein kinase C inhibitor, staurosporine. , 1992, European journal of pharmacology.

[23]  T. Tsuruo,et al.  Reversal mechanism of multidrug resistance by verapamil: direct binding of verapamil to P-glycoprotein on specific sites and transport of verapamil outward across the plasma membrane of K562/ADM cells. , 1989, Cancer research.

[24]  K. L. Peterson,et al.  Inhibitors of Ca2+ channels, calmodulin and protein kinases prevent A23187 and other inductions of metallothionein mRNA in EC3 rat hepatoma cells. , 1993, Life sciences.

[25]  T. Chambers,et al.  Protein kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cells. , 1990, The Journal of biological chemistry.

[26]  D. Reis,et al.  Agmatine: an endogenous clonidine-displacing substance in the brain. , 1994, Science.

[27]  C V Byus,et al.  Regulation of putrescine export in lipopolysaccharide or IFN-gamma-activated murine monocytic-leukemic RAW 264 cells. , 1994, Journal of immunology.

[28]  R. S. Gilbert,et al.  An ion-exchange chromatography procedure for the isolation and concentration of basic amino acids and polyamines from complex biological samples prior to high-performance liquid chromatography. , 1991, Analytical biochemistry.

[29]  R. Minchin,et al.  Evidence for the existence of distinct transporters for the polyamines putrescine and spermidine in B16 melanoma cells. , 1991, European journal of biochemistry.

[30]  M. Debenedette,et al.  Expression of polyamine transporter activity during B lymphocyte cell cycle progression. , 1993, Journal of immunology.

[31]  W. Catterall,et al.  Receptor sites for Ca2+ channel antagonists. , 1992, Trends in pharmacological sciences.

[32]  S. Bates,et al.  Differential modulation of P-glycoprotein transport by protein kinase inhibition. , 1993, Biochemistry.

[33]  K. Williams,et al.  Modulation of the NMDA receptor by polyamines. , 1991, Life sciences.

[34]  A. Galston,et al.  7 – Polyamine Metabolism , 1990 .

[35]  A. Pegg.  Recent advances in the biochemistry of polyamines in eukaryotes. , 1986, The Biochemical journal.

[36]  R. Fine,et al.  Phorbol esters induce multidrug resistance in human breast cancer cells. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[37]  T. Nguyen,et al.  Regulation of aldosterone biosynthesis by Na+/H+ antiport: relationships between intracellular pH and angiotensin II. , 1989, Endocrinology.

[38]  N. Seiler.  Functions of polyamine acetylation. , 1987, Canadian journal of physiology and pharmacology.

[39]  B. Chauffert,et al.  Sufficient levels of quinine in the serum circumvent the multidrug resistance of the human leukemic cell line K562/ADM , 1991, Cancer.

[40]  C V Byus,et al.  Selective putrescine export is regulated by insulin and ornithine in Reuber H35 hepatoma cells. , 1994, Biochimica et biophysica acta.

[41]  C V Byus,et al.  Biosynthesis and selective export of 1,5-diaminopentane (cadaverine) in mycoplasma-free cultured mammalian cells. , 1994, The Journal of biological chemistry.

[42]  K. Kashiwagi,et al.  Excretion of putrescine by the putrescine-ornithine antiporter encoded by the potE gene of Escherichia coli. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[43]  S. Yamamoto,et al.  Differential inhibition by staurosporine, a potent protein kinase C inhibitor, of 12-O-tetradecanoylphorbol-13-acetate-caused skin tumor promotion, epidermal ornithine decarboxylase induction, hyperplasia and inflammation. , 1989, Carcinogenesis.