ライフサイエンスコーパス: NCX1

1 NCX1 blockade impacted on CaMKII signalling to down-regu

2 NCX1 current densities were decreased by > 80 and 40%, r

3 NCX1 current measurements indicate that PLM decreased NC

4 NCX1 full-length protein and a 75-kDa NCX1 fragment alon

5 NCX1 is a Na(+)/Ca(2+) exchanger, which is believed to p

6 NCX1 is implicated in the pathogenesis of heart failure

7 NCX1 is made up of N-terminal domain consisting of the f

8 NCX1 palmitoylation is governed by a distal secondary st

9 NCX1, the protein encoded by SLC8A1, was expressed in sp

10 reverse mode of NCX subtypes 3 (NCX3) and 1 (NCX1), respectively, on the occurrence of acute seizures

11 ger 1 (NHE1) and sodium-calcium exchanger 1 (NCX1) are membrane proteins located on the bleb membrane

12 sodium (Na(+))-calcium (Ca(2+)) exchanger 1 (NCX1) is an important regulator of intracellular Ca(2+)

13 sodium (Na(+))-calcium (Ca(2+)) exchanger 1 (NCX1) is central to the maintenance of normal Ca(2+) hom

14 The sodium-calcium exchanger 1 (NCX1) is predominantly expressed in the heart and is imp

15 The Na(+)-Ca(2+) exchanger 1 (NCX1) is reduced in stroke by the RE1-silencing transcri

16 ice expressing the Na(+)-Ca(2+) exchanger 1 (NCX1) to model its identified augmentation during muscul

17 rated earlier that Na(+)/Ca(2+) exchanger 1 (NCX1), a major calcium exporter in renal epithelial cell

18 forms 2 and 3, and Na(+)/Ca(2+)-exchanger 1 (NCX1).

19 Na(+)/Ca(2+) exchanger-1 (NCX1) is a major calcium extrusion mechanism in renal ep

20 Na(+)/Ca(2+) exchanger type 1 (NCX1), a key PM Ca(2+) transporter, was immunoprecipitat

21 pA pF-1) and frog/dog (0.6 +/- 0.1 pA pF-1) NCX1, but less so in those expressing S374G frog NCX1 (0

22 In addition to transporting Ca(2+), NCX1.1 activity is also strongly regulated by Ca(2+) bin

23 This GlyT2.PMCA2,3.NCX1 complex is found in lipid raft subdomains where Gly

24 de that direct effects of PIP(2) to activate NCX1 can be strongly modulated by opposing mechanisms in

25 ion, which would favor greater NKA activity, NCX1 activity, and Ca2+ removal.

26 etween cysteines at position 122 in adjacent NCX1 proteins.

27 l side of Cys-739 in YFP-NCX1 did not affect NCX1 palmitoylation, with the exception of the rare huma

28 Our finding of an NCX1 macromolecular complex in heart suggests how NCX1 r

29 ylation is attributed to the existence of an NCX1 macromolecular complex.

30 eatment in mice leads to the formation of an NCX1-p38 complex.

31 ed and coimmunoprecipitated with GAP-43, and NCX1 silencing prevented NGF-induced effects on GAP-43 e

32 indings reveal a novel function for NHE1 and NCX1 in membrane blebbing and permeability, and establis

33 =1muM, while dually inhibiting both NHE1 and NCX1.1 at >/=20muM.

34 00 are more selective inhibitors of NHE1 and NCX1.1 than amiloride or DCB, respectively.

35 rescence studies that both exogenous PLM and NCX1 co-localized at the plasma membrane.

36 Co-expression of PLM and NCX1 inhibited NCX1 current (both modes).

37 em that is devoid of both endogenous PLM and NCX1, we first demonstrated by confocal immunofluorescen

38 s that are devoid of both endogenous PLM and NCX1, we first demonstrated that the exogenous NCX1 curr

39 between the transmembrane domains of PLM and NCX1.

40 cells that are devoid of endogenous PLM and NCX1.

41 protein-protein interaction between PLM and NCX1.

42 luorescent protein-NCX1 fusion proteins, and NCX1 currents using whole-cell voltage clamping.

43 there is dense labeling of SERCA2, RyRs, and NCX1 in small-sized SANCs, thought to reside within the

44 e analysed effects of aortic banding because NCX1 currents do not mirror the increases of NCX1 messag

45 only when alpha1D was silenced, and blocking NCX1/3 increased cytosolic Ca(2+) concentration and cell

46 nefits and risks of therapeutically blocking NCX1 in heart failure and during ischemia-reperfusion ar

47 odium efflux (and calcium influx) induced by NCX1 in a reverse mode results in membrane bleb retracti

48 by compensating net Na(+) charge movement by NCX1.

49 nsistent with destabilization of E-cadherin, NCX1 knockdown cells showed an increase in beta-catenin

50 g myocytes and in myocytes expressing canine NCX1.1 with the 562-679 f-loop deletion (NCX-(Delta562-6

51 CBD1, the primary Ca(2+) sensor from canine NCX1, but not the Ca(2+)-free form, has been reported, a

52 with a unique splice variant of frog cardiac NCX1 where insertion of an extra exon completes the codi

53 the electrogenic sarcolemma membrane cardiac NCX1 can act as a regulator of "activity-dependent signa

54 ovel endogenous protein inhibitor of cardiac NCX1.

55 he Ca(2+)-activated protease calpain cleaves NCX1.

56 In conclusion, NCX1 is a substrate-specifying PP1c regulator protein, i

57 rack NCX1 and study changes in conformation, NCX1 was tagged with derivatives of green fluorescent pr

58 neurons and glia PM microdomains containing NCX1 and Na(+) pumps with alpha2 or alpha3 subunits form

59 NCX1 large intracellular loop that controls NCX1 palmitoylation.

60 exhibits robust positive labeling for cRyR, NCX1, and SERCA2.

61 o cysteine on the backbone of a cysteineless NCX1.

62 ent measurements indicate that PLM decreased NCX1 current only when the split exchangers contained re

63 rog NCX1, and absent in cells expressing dog NCX1.

64 frog NCX1 (frog NCX1); (3) chimeric frog-dog NCX1 incorporating the completed P-loop from the frog NC

65 also downregulated in the frog and frog/dog NCX1 and to a smaller and transient extent in S374G frog

66 cAMP) suppressed INa-Ca of frog and frog/dog NCX1 by 60-80 %.

67 og NCX1 into the dog NCX1 sequence (frog/dog NCX1); and (4) a mutated frog NCX1 where a putative prot

68 lls) expressing: (1) wild-type dog NCX1 (dog NCX1); (2) wild-type frog NCX1 (frog NCX1); (3) chimeric

69 leted P-loop from the frog NCX1 into the dog NCX1 sequence (frog/dog NCX1); and (4) a mutated frog NC

70 y (HEK) cells) expressing: (1) wild-type dog NCX1 (dog NCX1); (2) wild-type frog NCX1 (frog NCX1); (3

71 SLC8A1), a sodium/calcium exchanger encoding NCX1, were validated in an independent Japanese genome-w

72 H9c2 expresses EAATs but lacks endogenous NCX1 expression.

73 Engineering NCX1-Met(369) into a tobacco etch virus protease cleavag

74 this isoform of NKA preferentially enhances NCX1 activity compared with NKA-alpha1.

75 Cardiac Na(+)-Ca(2+) exchange (NCX1) inactivates in excised membrane patches when cytop

76 The Na(+)/Ca(2+) exchanger NCX1 was found to interact with Ano6 in a two-hybrid spl

77 ID2 and the cardiac sodium-calcium exchanger NCX1 in the proximal CCS, where GATA6 transactivates bot

78 ions induced by the sodium/calcium exchanger NCX1/3 working in its reverse mode.

79 d precise control of Na(+)/Ca(2+) exchanger (NCX1) activity is essential in the maintenance of beat-t

80 al expression of the Na(+)-Ca(2+) exchanger (NCX1) and L-type Ca(2+) channel (LTCC).

81 egarding the cardiac Na(+)/Ca(2+) exchanger (NCX1) and Na(+)/K(+) pump and the controversies that sti

82 The Na(+)/Ca(2+) exchanger (NCX1) is a plasma membrane protein important in regulati

83 The cardiac Na (+)-Ca (2+) exchanger (NCX1) is modeled to contain nine transmembrane segments

84 These activate the Na(+)/Ca(2+) exchanger (NCX1) to generate inward current and membrane excitation

85 ivity of the cardiac Na(+)-Ca(2+) exchanger (NCX1.1) is allosterically regulated by Ca(2+), which bin

86 The cardiac Na(+)/Ca(2+) exchanger (NCX1.1) serves as the primary means of Ca(2+) extrusion

87 mman (PLM), and the cardiac Na/Ca exchanger (NCX1) show greater surface membrane localization, and ME

88 The cardiac sarcolemmal Na+-Ca2+ exchanger (NCX1) influences cardiac contractility by extruding Ca2+

89 The cardiac Na+-Ca2+ exchanger (NCX1) is a membrane protein that extrudes Ca2+ from cell

90 The cardiac Na+-Ca2+ exchanger (NCX1) is one of the major sarcolemmal Ca2+ transporters

91 The Na+-Ca2+ exchanger (NCX1) is up-regulated in hypertrophy and is often found

92 racellular space via the Na+/Ca2+ exchanger (NCX1) or sequester it into the sarcoplasmic reticulum, v

93 lators, inhibits cardiac Na+/Ca2+ exchanger (NCX1).

94 in, inhibits the cardiac Na+/Ca2+ exchanger (NCX1).

95 isoform 1.1 of the sodium-calcium exchanger (NCX1.1), which increases [Ca(2+)](i) and initiates gliom

96 The mammalian Na(+)/Ca(2+) exchanger, NCX1.1, serves as the main mechanism for Ca(2+) efflux a

97 specific knockout of the Na+-Ca2+ exchanger, NCX1.

98 fraction of cardiac Na(+)-Ca(2+) exchangers (NCX1) in the surface membrane of cells (F(surf)).

99 X1, we first demonstrated that the exogenous NCX1 current (I(NaCa)) was increased by phorbol 12-myris

100 An extracellular NCX1 disulphide bond is rapidly reduced by tris(2-carbox

101 duced Na(+) clearance rates that would favor NCX1 reverse-mode operation, showed exacerbated disease

102 oaches suggest that PIP(2) synthesis favours NCX1 internalization, that NCX1 internalization is proba

103 aired palmitoylation of both YFP-NCX1 and FL-NCX1.

104 human polymorphism S738F, which enhanced FL-NCX1 palmitoylation, and D741A, which modestly reduced i

105 ngle amino acid changes around Cys-739 in FL-NCX1 and deletions on the N-terminal side of Cys-739 in

106 Full-length NCX1 (FL-NCX1) and a YFP fusion protein of the NCX1 large intrace

107 c face of the helix significantly reduced FL-NCX1 palmitoylation.

108 present an expanded helix-packing model for NCX1.

109 early heart and an essential early role for NCX1 in establishing SACOs through to the initiation of

110 ellular loop is necessary and sufficient for NCX1 palmitoylation.

111 ative PKA site, present in both dog and frog NCX1, might also be critical in the cAMP-mediated regula

112 ence (frog/dog NCX1); and (4) a mutated frog NCX1 where a putative protein kinase A (PKA) site was di

113 X1 (dog NCX1); (2) wild-type frog NCX1 (frog NCX1); (3) chimeric frog-dog NCX1 incorporating the comp

114 , but less so in those expressing S374G frog NCX1 (0.3 +/- 0.1 pA pF-1).

115 ngle serine residue with glycine (S374G frog NCX1).

116 and transient in cells expressing S374G frog NCX1, and absent in cells expressing dog NCX1.

117 a smaller and transient extent in S374G frog NCX1.

118 rporating the completed P-loop from the frog NCX1 into the dog NCX1 sequence (frog/dog NCX1); and (4)

119 he presence of the P-loop domain of the frog NCX1, and provide evidence that the putative PKA site, p

120 type dog NCX1 (dog NCX1); (2) wild-type frog NCX1 (frog NCX1); (3) chimeric frog-dog NCX1 incorporati

121 ion, and Ca(2+) increase unless a functional NCX1 was introduced in H9c2 cells by stable transfection

122 nd hypothesized that a direct and functional NCX1-PP1 interaction is a prerequisite for pSer-68-PLM d

123 macromolecular complex in heart suggests how NCX1 regulation is achieved in heart and other cells.

124 ctrin and IP(3)R-1 all co-immunoprecipitated NCX1.

125 forms, but not alpha1, co-immunoprecipitated NCX1; the antibodies to alpha1 did, however, co-immunopr

126 n the first Ca(2+) binding domain (CBD) 1 in NCX1.

127 The palmitoylated cysteine in NCX1 is found in all vertebrate and some invertebrate NC

128 chored to the first Ca(2+) binding domain in NCX1, whereas the calpain catalytic region bound to the

129 c region bound to the catenin-like domain in NCX1.

130 tical Ca(2+) binding sites as those found in NCX1 and that the partial Ca(2+) occupancy and apoform s

131 KO) of Na,K-beta had a specific reduction in NCX1 protein and were ouabain-insensitive.

132 lation and GAP-43 protein expression rise in NCX1.4 overexpressing cells.

133 ) ions in NCX_Mj, play a fundamental role in NCX1.1.

134 Thus, G-protein signalling may increase NCX1 currents by destabilizing membrane cytoskeleton-PIP

135 Na(+) channel inhibitor that should increase NCX1 forward-mode operation, reduced muscular pathology.

136 eta-catenin, GATA4, NFATc3/c4, and increased NCX1, nuclear DYKR1A, and Pur alpha/beta protein.

137 E2 incubation (24 h) increased NCX1 (50%) and INCX ( approximately 3-fold at 60 mV) in

138 athophysiological conditions where increased NCX1 activity contributes to cardiac dysfunction.

139 rotective effect during stroke and increases NCX1 and NCX2 activities.

140 ssion of three potassium (K(+))-independent (NCX1, 2, 3) and three K(+)-dependent (NCKX1, 2, 3) Na(+)

141 have observed here following calpain-induced NCX1 cleavage, might be beneficial in pathophysiological

142 that specific cleavage at Met(369) inhibited NCX1 activity (both forward and reverse mode).

143 Co-expression of PLM and NCX1 inhibited NCX1 current (both modes).

144 rylated phospholemman (pSer-68-PLM) inhibits NCX1 activity.

145 Interestingly, NCX1/3 regulated membrane potential of HCT116 cells only

146 expression of its neuronal splicing isoform, NCX1.4, even in the absence of NGF, induced an increase

147 hereas mRNA encoding all three NCX isoforms (NCX1-3) was detected in putative nociceptive cutaneous n

148 three Na(+)/Ca(2+) exchanger (NCX) isoforms, NCX1, NCX2, and NCX3, worsens ischemic brain damage.

149 NCX1 full-length protein and a 75-kDa NCX1 fragment along with calpain were up-regulated in ao

150 Full-length NCX1 (FL-NCX1) and a YFP fusion protein of the NCX1 larg

151 lobulin-like structure, similar to mammalian NCX1-CBD2, but the predicted Ca(2+) interaction region o

152 by whole-cell patch clamp studies to measure NCX1 current density and radiotracer flux assays to asse

153 We measured NCX1 palmitoylation using resin-assisted capture, the su

154 ough a predicted enhancement in forward-mode NCX1 operation that reduces Ca(2+) levels.

155 Based on the current topological model, NCX1 consists of nine transmembrane segments (TMSs).

156 Moreover, NCX1 colocalized and coimmunoprecipitated with GAP-43, a

157 were transfected with cDNAs encoding mutant NCX1 proteins.

158 Moreover, in primary cortical neurons, NCX1 silencing prevented Akt phosphorylation, GAP-43 and

159 ween CRMP2 and NMDAR as well as NCX3 but not NCX1.

160 ues within transmembrane segments 2 and 7 of NCX1.1 (cardiac isoform) to cysteines, allowing us to in

161 insights into the TMS packing arrangement of NCX1, we performed cysteine cross-linking experiments.

162 independent, and that significant changes of NCX1 surface expression occur physiologically and pathol

163 sequences of calpain binding and cleavage of NCX1 in the heart.

164 ere separated (mimicking calpain cleavage of NCX1).

165 Submembrane colocalization of NCX1 and cardiac RyR (cRyR) in all SANCs exceeds that in

166 emical staining showed the colocalization of NCX1 protein with mAKAP and PKA-RI proteins in cardiomyo

167 s to identify the structural determinants of NCX1 palmitoylation.

168 is, three specific molecular determinants of NCX1 responsible for neurounina-1 activity were identifi

169 PLM associated with the N-terminal domain of NCX1 when it contained intracellular loop residues 218-3

170 he N- or C-terminal transmembrane domains of NCX1, was associated with PLM.

171 tified from Western blots as the fraction of NCX1 that migrates at a higher molecular weight.

172 The function of NCX1 depends on subcellular ("local") factors, the phosp

173 Our findings demonstrate the importance of NCX1 for bone mineralization and explain why deletion of

174 NCX1 currents do not mirror the increases of NCX1 message and protein that occur in this model.

175 -cadherin to the cell surface independent of NCX1 ion transport activity.

176 ation of PLM with NCX1 was the inhibition of NCX1 activity, as demonstrated by whole-cell patch clamp

177 Inhibition of NCX1 activity, such as we have observed here following c

178 Pharmacological inhibition of NCX1 and LTCC revealed rapid development of Ca(2+) handl

179 In parallel, inhibition of NCX1 by KB-R7943 in Madin-Darby canine kidney cells, LLC

180 physiological significance of inhibition of NCX1 by phosphorylated PLM was evaluated in PLM-knock-ou

181 Inhibition of NCX1 by PLM was specific, because a single mutation of s

182 resulted in a complete loss of inhibition of NCX1 current, although association of the PLM mutant wit

183 However, inhibition of NCX1 enhances cell blebbing; cells become swollen becaus

184 s suggest that beta-adrenergic inhibition of NCX1, as reported for frog, but not mammalian hearts, ma

185 Knockdown of NCX1 (the main NCX isoform in HUVECs) by siRNA confirmed

186 Taken together, knockdown of NCX1 in Madin-Darby canine kidney cells alters epithelia

187 Knockdown of NCX1 in Madin-Darby canine kidney cells induced fibrobla

188 denal epithelial cell line, and knockdown of NCX1 proteins with a specific siRNA greatly decreased th

189 Labeling of NCX1 is also similar among SANCs of all sizes and exceed

190 g heart and suggests that relative levels of NCX1 and SERCA2 function are essential for normal develo

191 wed exacerbated disease in the hind limbs of NCX1 TG mice, similar to treatment with the Na(+)-K(+) A

192 not influence trafficking or localization of NCX1 to surface membranes, nor does it strongly affect t

193 ate of NCX1, and the subcellular location of NCX1 within the cell.

194 7, or 548 of the large intracellular loop of NCX1 located between transmembrane segments 5 and 6.

195 or YFP into the large intracellular loop of NCX1 protein does not impair exchanger properties.

196 arious segments of the intracellular loop of NCX1 suggest that PLM bound to residues 218-371 and 508-

197 The cytosolic loop of NCX1, containing the K(I/V)FF motif, had no effect on PP

198 LM interacted with the intracellular loop of NCX1, most likely at residues 218-358.

199 sities were decreased by 78% with no loss of NCX1 expression.

200 s to selectively inhibit the forward mode of NCX1.1 at </=1muM, while dually inhibiting both NHE1 and

201 normal forward or reverse transport modes of NCX1.

202 nflux state due to reverse-mode operation of NCX1, which mediates disease.

203 we investigate the molecular organization of NCX1 within the cardiac myocyte.

204 Palmitoylation of NCX1 occurs in the Golgi and anchors the NCX1 large regu

205 FRET analysis indicates the proximity of NCX1 to plasma membrane phosphatidylinositol 4,5-bisphos

206 e replaced with the corresponding regions of NCX1.

207 of the P-loop in cAMP-mediated regulation of NCX1 we used four stably transfected human cell lines (a

208 Negative regulation of NCX1- mediated renal Ca(2+) absorption by IFNgamma may s

209 their possible involvement in regulation of NCX1.1 activity.

210 u-454 have no impact on Ca(2+) regulation of NCX1.1.

211 ed that calpain is an important regulator of NCX1 in response to pressure overload and aimed to ident

212 These data confirm the role of NCX1 activity in regulating renal epithelial cell migrat

213 ) pacemaker cells, but the potential role of NCX1 in determining heart rate in vivo is unknown.

214 addition, in neurons silenced with siRNA of NCX1 and subjected to oxygen and glucose deprivation (OG

215 ocal") factors, the phosphorylation state of NCX1, and the subcellular location of NCX1 within the ce

216 intracellular [Ca(2+)], or disabling RyRs or NCX1, markedly attenuates or abolishes spontaneous SANC

217 Outward NCX1 currents increase as expected.

218 As expected, outward NCX1 current (i.e. Ca(2+) influx) can be strongly inhibi

219 nificantly increased in cells overexpressing NCX1.4 as well as ER Ca(2+) content.

220 nteracted, whereas in neurons overexpressing NCX1 and subjected to ischemic preconditioning (PC+OGD/R

221 were similar, suggesting that physiological NCX1 expression is not required for determining resting

222 lular location of yellow fluorescent protein-NCX1 fusion proteins, and NCX1 currents using whole-cell

223 Rat NCX1 is substantially palmitoylated in all tissues exami

224 cells with Na,K-beta knockdown have reduced NCX1 protein and function accompanied by 2.1-fold increa

225 ymes are coordinately positioned to regulate NCX1 as has been found in diverse cells for a number of

226 P1c regulator protein, indirectly regulating NCX1 activity through pSer-68-PLM dephosphorylation.

227 Ca2+ from the cytosol in a manner requiring NCX1 activity.

228 Restoring NCX1 expression in beta-KD cells reduced migration rate

229 lly, muscle-specific deletion of the Slc8a1 (NCX1) gene diminished hind-limb pathology in Sgcd(-/-) m

230 substrate competitor peptide, and a specific NCX1-Met(369) antibody identified a novel calpain cleava

231 t here that mutation of the cardiac-specific NCX1 (NCX1h) gene causes embryonic lethal cardiac arrhyt

232 -0.4 are obtained for cell lines with stable NCX1 expression, 0.3 for neonatal myocytes and 0.6-0.8 f

233 Third, using BHK cells with stable NCX1 expression, we increased PIP(2) by transient expres

234 niques revealed that neurounina-1 stimulated NCX1 and NCX2 activities with an EC(50) in the picomolar

235 ation of untagged NCX1 with histidine-tagged NCX1.

236 used yellow fluorescent protein (YFP)-tagged NCX1 to redistribute diffusely into the cytoplasm within

237 Collectively, these results demonstrate that NCX1 expression is regulated by the Sp3/REST/HDAC1/HDAC2

238 m Ano6(-/-) and WT mice, we demonstrate that NCX1 requires Ano6 to efficiently translocate Ca(2+) out

239 molecular cross-link, we found evidence that NCX1 can form dimers.

240 We also provide evidence that NCX1 interacts with and anchors E-cadherin to the cell s

241 liminated in Ncx1(-/-) mice, indicating that NCX1 is important for fight or flight heart rate respons

242 synthesis favours NCX1 internalization, that NCX1 internalization is probably clathrin-independent, a

243 We show that NCX1 is dynamically phosphorylated by protein kinase A (

244 Furthermore, we show that NCX1 mRNA in rat olfactory mucosa is expressed as 8 alte

245 Collectively, these data show that NCX1 participates in neuronal differentiation through th

246 We now show that NCX1, like Cav1.2alpha, is greater at the base of female

247 rate and ERK1/2 activation, suggesting that NCX1 functions downstream of Na,K-beta in regulating cel

248 d immunohistochemical analysis suggests that NCX1 is redistributed away from the outer sarcolemma.

249 Here, we show for the first time that NCX1 mRNA and protein expression was down-regulated in W

250 The NCX1 (sodium-calcium exchanger) is up-regulated in human

251 The NCX1 transgene induced dystrophy-like disease in all hin

252 The NCX1-generated current (INa-Ca) was reliably measured in

253 of NCX1 occurs in the Golgi and anchors the NCX1 large regulatory intracellular loop to membranes.

254 lly, a short peptide fragment containing the NCX1-Met(369) cleavage site was modeled into the narrow

255 gly, during NGF-induced differentiation, the NCX1 protein level increased, NCX3 decreased, and NCX2 r

256 Cysteine 739 in the NCX1 large intracellular loop is necessary and sufficien

257 identified an amphipathic alpha-helix in the NCX1 large intracellular loop that controls NCX1 palmito

258 e residues that coordinate two Ca(2+) in the NCX1-CBD1 structure are neutralized by two Lys residues

259 domains as a result of Ca(2+) binding in the NCX1.1 isoform.

260 nstrate that prolonged administration of the NCX1 inhibitor KB-R7943 resulted in the up-regulation of

261 The myocardium of the NCX1 knockout mice undergoes a remarkable adaptation to

262 X1 (FL-NCX1) and a YFP fusion protein of the NCX1 large intracellular loop (YFP-NCX1) were expressed

263 The existence of the NCX1 macromolecular complex may also provide an explanat

264 Surprisingly, the NCX1 knockout mice live to adulthood with only modestly

265 over, immunohistochemistry revealed that the NCX1, 2, and 3 proteins are expressed in nearly all neur

266 alyze the mechanisms of PP1 targeting to the NCX1-pSer-68-PLM complex and hypothesized that a direct

267 Structural expression of these NCX1 constructs was confirmed using Western blot analysi

268 Of these, NCX1 and NCKX1 represent the most and least abundant mRN

269 me degree of reverse-mode Ca2+ entry through NCX1, as well as less efficient Ca2+ clearance.

270 To track NCX1 and study changes in conformation, NCX1 was tagged

271 ts the sodium-dependent calcium transporter (NCX1.1) much more potently than NHE1.

272 By trapping NCX1.1 in the inward-facing configuration, we have mappe

273 ilar biophysical properties to the wild type NCX1.

274 did not co-immunoprecipitate with wild-type NCX1 when co-expressed in HEK293 cells, confirming littl

275 In cells expressing wild-type NCX1.1, increasing concentrations of cytosolic Na(+) led

276 activation of NCX activity in the wild-type NCX1.1; indeed, at elevated [Na(+)], the D447V mutant be

277 that demonstrate the association of untagged NCX1 with histidine-tagged NCX1.

278 Thus, E2 upregulates NCX1 by a genomic mechanism mediated by ERs, and de novo

279 F(surf) remains less than 0.1 when NCX1 is expressed via transient transfections.

280 68-PLM in HEK293 cells was not observed when NCX1 was absent, when the K(I/V)FF motif was mutated, or

281 e effect is observed in the diaphragm, where NCX1 overexpression mildly protects from dystrophic dise

282 We investigated whether NCX1, NCX2, and NCX3 isoforms could play a relevant role

283 d PIP(2) were increased significantly, while NCX1 current densities were decreased by 78% with no los

284 regulatory enzymes are also associated with NCX1, including protein kinase C (PKC) and two serine/th

285 e demonstrate that Na,K-beta associates with NCX1 and regulates its localization to the cell surface.

286 PP1 also associates with NCX1; however, the molecular basis of this association i

287 When co-expressed with NCX1, PLM resulted in: (i) decreases in I(NaCa), (ii) at

288 l proteins did not co-immunoprecipitate with NCX1, including the Na(+) pump alpha1 isoform, PM Ca(2+)

289 fractionated, and co-immunoprecipitated with NCX1 in rat cardiomyocytes and left ventricle lysate.

290 fractionated, and co-immunoprecipitated with NCX1 in rat cardiomyocytes, left ventricle lysates, and

291 also selectively co-immunoprecipitated with NCX1, as did the ER proteins, Ca(2+) pump type 2 (SERCA

292 ndant PM proteins co-immunoprecipitated with NCX1, including the alpha2 and alpha3 isoforms of the Na

293 iac myocytes, PLM co-immunoprecipitated with NCX1.

294 although association of the PLM mutant with NCX1 was unaltered.

295 nsequences of direct association of PLM with NCX1 was the inhibition of NCX1 activity, as demonstrate

296 Moreover, co-expression of PLM with NCX1(F407P) (mutated K(I/V)FF motif) resulted in the cur

297 the C-terminal side of Cys-739 abolished YFP-NCX1 palmitoylation.

298 prolines impaired palmitoylation of both YFP-NCX1 and FL-NCX1.

299 ons on the N-terminal side of Cys-739 in YFP-NCX1 did not affect NCX1 palmitoylation, with the except

300 in of the NCX1 large intracellular loop (YFP-NCX1) were expressed in HEK cells.