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J Physiol
2003 May 01;548Pt 3:677-89. doi: 10.1113/jphysiol.2002.036426.
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Molecular determinants of cAMP-mediated regulation of the Na+-Ca2+ exchanger expressed in human cell lines.
He LP
,
Cleemann L
,
Soldatov NM
,
Morad M
.
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The cardiac Na+-Ca2+ exchanger (NCX1) is one of the major sarcolemmal Ca2+ transporters of cardiomyocytes. Structure-function studies suggest that beta-adrenergic inhibition of NCX1, as reported for frog, but not mammalian hearts, may be associated with a unique splice variant of frog cardiac NCX1 where insertion of an extra exon completes the coding of a nucleotide binding P-loop. To test the involvement of the P-loop in cAMP-mediated regulation of NCX1 we used four stably transfected human cell lines (a previously established line of baby hamster kidney (BHK) cells and three new lines of human embryonic kidney (HEK) cells) expressing: (1) wild-type dog NCX1 (dog NCX1); (2) wild-type frog NCX1 (frog NCX1); (3) chimeric frog-dog NCX1 incorporating the completed P-loop from the frog NCX1 into the dog NCX1 sequence (frog/dog NCX1); and (4) a mutated frog NCX1 where a putative protein kinase A (PKA) site was disrupted by substitution of a single serine residue with glycine (S374G frog NCX1). Structural expression of these NCX1 constructs was confirmed using Western blot analysis of extracted proteins and immunofluorescence imaging. The NCX1-generated current (INa-Ca) was reliably measured in cells expressing dog (2.0 +/- 0.15 pA pF-1), frog (0.6 +/- 0.1 pA pF-1) and frog/dog (0.6 +/- 0.1 pA pF-1) NCX1, but less so in those expressing S374G frog NCX1 (0.3 +/- 0.1 pA pF-1). Addition of 100 microM 8-bromoadenosine 3',5' cyclic monophosphate (8-Br-cAMP) suppressed INa-Ca of frog and frog/dog NCX1 by 60-80 %. The suppression of INa-Ca was smaller and transient in cells expressing S374G frog NCX1, and absent in cells expressing dog NCX1. Intracellular Ca2+ (Ca2+i)-transients, activated by rapid withdrawal of Na+, were also downregulated in the frog and frog/dog NCX1 and to a smaller and transient extent in S374G frog NCX1. Our findings suggest that the suppressive effect of beta-adrenergic agonists requires the presence of the P-loop domain of the frog NCX1, and provide evidence that the putative PKA site, present in both dog and frog NCX1, might also be critical in the cAMP-mediated regulation of the exchanger.
Baker,
The influence of calcium on sodium efflux in squid axons.
1969, Pubmed
Baker,
The influence of calcium on sodium efflux in squid axons.
1969,
Pubmed
Ballard,
Stimulation of the Na+/Ca2+ exchanger by phenylephrine, angiotensin II and endothelin 1.
1996,
Pubmed
Chen,
A Ca2+-dependent tryptic cleavage site and a protein kinase A phosphorylation site are present in the Ca2+ regulatory domain of scallop muscle Na+-Ca2+ exchanger.
2000,
Pubmed
Cleemann,
Role of Ca2+ channel in cardiac excitation-contraction coupling in the rat: evidence from Ca2+ transients and contraction.
1991,
Pubmed
DiPolo,
Calcium influx in internally dialyzed squid giant axons.
1979,
Pubmed
Fabiato,
Calcium-induced release of calcium from the sarcoplasmic reticulum of skinned cells from adult human, dog, cat, rabbit, rat, and frog hearts and from fetal and new-born rat ventricles.
1978,
Pubmed
Fan,
Regulation of cardiac sodium-calcium exchanger by beta-adrenergic agonists.
1996,
Pubmed
Fischmeister,
Mechanism of action of acetylcholine on calcium current in single cells from frog ventricle.
1986,
Pubmed
Frank,
Distribution of the Na(+)-Ca2+ exchange protein in mammalian cardiac myocytes: an immunofluorescence and immunocolloidal gold-labeling study.
1992,
Pubmed
Goaillard,
Simultaneous measurements of intracellular cAMP and L-type Ca2+ current in single frog ventricular myocytes.
2001,
Pubmed
Hamill,
Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
1981,
Pubmed
Han,
Contribution of Na(+)-Ca2+ exchange to stimulation of transient inward current by isoproterenol in rabbit cardiac Purkinje fibers.
1995,
Pubmed
Hilgemann,
Regulation and deregulation of cardiac Na(+)-Ca2+ exchange in giant excised sarcolemmal membrane patches.
1990,
Pubmed
Hinde,
Inhibition of Na/Ca exchange by external Ni in guinea-pig ventricular myocytes at 37 degrees C, dialysed internally with cAMP-free and cAMP-containing solutions.
1999,
Pubmed
Iwamoto,
A novel isothiourea derivative selectively inhibits the reverse mode of Na+/Ca2+ exchange in cells expressing NCX1.
1996,
Pubmed
Iwamoto,
Protein kinase C-dependent regulation of Na+/Ca2+ exchanger isoforms NCX1 and NCX3 does not require their direct phosphorylation.
1998,
Pubmed
Iwata,
A new splicing variant in the frog heart sarcolemmal Na-Ca exchanger creates a putative ATP-binding site.
1996,
Pubmed
,
Xenbase
Kavaler,
Paradoxical effects of epinephrine on excitation-contraction coupling in cardiac muscle.
1966,
Pubmed
Kim,
Cloning of the mouse cardiac Na(+)-Ca2+ exchanger and functional expression in Xenopus oocytes.
1996,
Pubmed
,
Xenbase
Kofuji,
Na/Ca exchanger isoforms expressed in kidney.
1993,
Pubmed
Komuro,
Molecular cloning and characterization of the human cardiac Na+/Ca2+ exchanger cDNA.
1992,
Pubmed
,
Xenbase
Kraev,
Molecular biological studies of the cardiac sodium-calcium exchanger.
1996,
Pubmed
Li,
Identification of a peptide inhibitor of the cardiac sarcolemmal Na(+)-Ca2+ exchanger.
1991,
Pubmed
Linck,
Functional comparison of the three isoforms of the Na+/Ca2+ exchanger (NCX1, NCX2, NCX3).
1998,
Pubmed
Low,
Cloning of the rat heart Na(+)-Ca2+ exchanger and its functional expression in HeLa cells.
1993,
Pubmed
Main,
Changes in subsarcolemmal sodium concentration measured by Na-Ca exchanger activity during Na-pump inhibition and beta-adrenergic stimulation in guinea-pig ventricular myocytes.
1997,
Pubmed
Manolopoulos,
Cyclic strain and forskolin differentially induce cAMP production in phenotypically diverse endothelial cells.
1993,
Pubmed
Martel,
The sensitivity of the human Kv1.3 (hKv1.3) lymphocyte K+ channel to regulation by PKA and PKC is partially lost in HEK 293 host cells.
1998,
Pubmed
Matsuoka,
Initial localization of regulatory regions of the cardiac sarcolemmal Na(+)-Ca2+ exchanger.
1993,
Pubmed
,
Xenbase
Matsuoka,
Regulation of the cardiac Na(+)-Ca2+ exchanger by Ca2+. Mutational analysis of the Ca(2+)-binding domain.
1995,
Pubmed
,
Xenbase
Maylie,
Excitation-contraction coupling in neonatal and adult myocardium of cat.
1982,
Pubmed
Menick,
The exchanger and cardiac hypertrophy.
1996,
Pubmed
Morad,
Contracture and catecholamines in mammalian myocardium.
1969,
Pubmed
Morad,
Relaxing effects of catecholamines on mammalian heart.
1972,
Pubmed
Morad,
The inotropic actions of adrenaline on frog ventricular muscle: relaxing versus potentiating effects.
1981,
Pubmed
Nicoll,
Cloning of a third mammalian Na+-Ca2+ exchanger, NCX3.
1996,
Pubmed
,
Xenbase
Nicoll,
Molecular cloning and functional expression of the cardiac sarcolemmal Na(+)-Ca2+ exchanger.
1990,
Pubmed
,
Xenbase
Nicoll,
Molecular studies of the cardiac sarcolemmal sodium-calcium exchanger.
1991,
Pubmed
Ottolia,
Split Na+-Ca2+ exchangers. Implications for function and expression.
2001,
Pubmed
,
Xenbase
Pabbathi,
Comparison of Na(+)/Ca(2+) exchanger current and of its response to isoproterenol between acutely isolated and short-term cultured adult ventricular myocytes.
2002,
Pubmed
Page,
Structures of physiological interest in the frog heart ventricle.
1972,
Pubmed
Perchenet,
Stimulation of Na/Ca exchange by the beta-adrenergic/protein kinase A pathway in guinea-pig ventricular myocytes at 37 degrees C.
2000,
Pubmed
Philipson,
Purification of the cardiac Na+-Ca2+ exchange protein.
1988,
Pubmed
Quednau,
Tissue specificity and alternative splicing of the Na+/Ca2+ exchanger isoforms NCX1, NCX2, and NCX3 in rat.
1997,
Pubmed
Reuter,
The Na+-Ca2+ exchanger is essential for the action of cardiac glycosides.
2002,
Pubmed
Reuter,
The dependence of calcium efflux from cardiac muscle on temperature and external ion composition.
1968,
Pubmed
Ruknudin,
Functional differences between cardiac and renal isoforms of the rat Na+-Ca2+ exchanger NCX1 expressed in Xenopus oocytes.
2000,
Pubmed
,
Xenbase
Saraste,
The P-loop--a common motif in ATP- and GTP-binding proteins.
1990,
Pubmed
Shuba,
A novel molecular determinant for cAMP-dependent regulation of the frog heart Na+-Ca2+ exchanger.
1998,
Pubmed
,
Xenbase
Smith,
Regulation of expression of sodium-calcium exchanger and plasma membrane calcium ATPase by protein kinases, glucocorticoids, and growth factors.
1996,
Pubmed
Vassort,
Membrane potential and slow inward current dependence of frog cardiac mechanical activity.
1972,
Pubmed
Vilsen,
Deduced amino acid sequence and E1-E2 equilibrium of the sarcoplasmic reticulum Ca(2+)-ATPase of frog skeletal muscle. Comparison with the Ca(2+)-ATPase of rabbit fast twitch muscle.
1992,
Pubmed
Watano,
A novel antagonist, No. 7943, of the Na+/Ca2+ exchange current in guinea-pig cardiac ventricular cells.
1996,
Pubmed
Woo,
Bimodal regulation of Na(+)--Ca(2+) exchanger by beta-adrenergic signaling pathway in shark ventricular myocytes.
2001,
Pubmed
Xue,
Cloning, expression, and characterization of the trout cardiac Na(+)/Ca(2+) exchanger.
1999,
Pubmed
,
Xenbase
