ライフサイエンスコーパス: GC-A

1 GC-A activity, but not protein, was increased in heart a

2 GC-A immunoprecipitates did not contain detectable amoun

3 GC-A was phosphorylated in the presence of [gamma32P]ATP

4 A GC-A mutant containing Glu substitutions for all seven c

5 dent phosphorylation sites is required for a GC-A conformation capable of transmitting the hormone bi

6 ad scaffold resulted in the development of a GC-A PAM, MCUF-651, which enhanced ANP-mediated cGMP gen

7 retic peptide (BNP) is a guanylyl cyclase A (GC-A) agonist.

8 etic peptide (ANP) binds guanylyl cyclase-A (GC-A) and natriuretic peptide receptor-C (NPR-C).

9 hrough disruption of the guanylyl cyclase-A (GC-A) natriuretic peptide receptor gene.

10 , via their cGMP-forming guanylyl cyclase-A (GC-A) receptor and cGMP-dependent kinase I (cGKI), stren

11 The guanylyl cyclase-A (GC-A) receptor is known to require both atrial natriuret

12 ic peptide (ANP) via its guanylyl cyclase-A (GC-A) receptor participates in regulation of arterial bl

13 Disruption of guanylyl cyclase-A (GC-A) results in mice displaying an elevated blood press

14 Guanylyl cyclase-A (GC-A) signaling, a natriuretic peptide receptor, exerts

15 rine output, and second, guanylyl cyclase-A (GC-A), the primary signaling receptor for BNP, is down-r

16 extracellular domain of guanylyl cyclase-A (GC-A), the receptor for atrial natriuretic peptide, five

17 Guanylyl cyclase-A (GC-A), the transmembrane cGMP-producing receptor shared

18 peptide (ANP) receptor [guanylyl cyclase-A (GC-A)] gene yields mice with a salt-resistant form of hy

19 stantly related guanylate cyclase isoform A (GC-A) gene shows the most divergence in the extracellula

20 yl cyclase-A/natriuretic peptide receptor-A (GC-A/NPRA) and produces the intracellular second messeng

21 isms of GC-A/natriuretic peptide receptor-A (GC-A/NPRA) gene (Npr1) expression in vivo.

22 ylyl cyclase/natriuretic peptide receptor-A (GC-A/NPRA) gene-targeted mice.

23 ylyl cyclase/natriuretic peptide receptor-A (GC-A/NPRA), which plays a critical role in reduction of

24 ylyl cyclase/natriuretic peptide receptor-A, GC-A/NPRA) gene-knockout (0-copy; Npr1(-/-) ), 2-copy (N

25 Natriuretic peptide receptor A (NPR-A/GC-A) and B (NPR-B/GC-B) are members of the transmembran

26 Guanylyl cyclase-A (NPR-A; GC-A) is the major and possibly the only receptor for at

27 fide ring structure of human BNP to activate GC-A and guanylyl cyclase-B (GC-B), which is not reduced

28 , neither staurosporine nor Go6976 activated GC-A or GC-B.

29 Because ATP allosterically activates GC-A and GC-B, we investigated how ATP binding to the PK

30 Although GC-A is highly phosphorylated, and dephosphorylation ina

31 sed and decreased potency for human GC-B and GC-A, respectively.

32 A physical association between hsp90 and GC-A was demonstrated in coimmunoprecipitation experimen

33 xamined the interaction between p38 MAPK and GC-A signaling.

34 ignificantly different between wild-type and GC-A null mice on standard rodent chow (0.7% NaCl) or a

35 were markedly elevated in both wild-type and GC-A-null mice.

36 ecreased aldosterone in male GC-A(WT/WT) and GC-A(8E/8E) mice at 15 min, but only GC-A(8E/8E) mice ha

37 racellular guanylyl cyclase domains, such as GC-A and GC-B, also known as Npr1 and Npr2, respectively

38 P, membrane-bound guanylyl cyclases A and B (GC-A and GC-B), mediate the effects of NPs via the gener

39 Complexes between GC-A and hsp90 contained the co-chaperone p50(cdc37), ty

40 Defects in the BNP/GC-A/cGMP pathway may play a role in arteriopathies in w

41 l-bnp also bound GC-A 4.5-fold less tightly than wild type.

42 tify in vivo processes that are regulated by GC-A phosphorylation, we substituted glutamates for know

43 to the accumulation of complexes containing GC-A and heat shock protein 70 (hsp70).

44 Activation of particulate guanylate cyclase (GC-A) by ANP leads to a substantial, dose-dependent, rap

45 ude that increased phosphorylation-dependent GC-A activity decreases cardiac ERK activity, which resu

46 on of albumin clearances in hypervolaemic EC GC-A KO mice with normovolaemic littermates demonstrated

47 loxed GC-A x tie2-Cre: endothelial cell (EC) GC-A knockout (KO)); and (iii) control littermates (flox

48 attenuated in knockout mice with endothelial GC-A deletion and unaltered in knockout mice with endoth

49 tion of GC-A (knockout mice with endothelial GC-A deletion) or cGKI (knockout mice with endothelial c

50 Here, we evaluated GC-A and NPR-C internalization using traditional and nov

51 Although HeLa cells endogenously express GC-A, (125)I-ANP binding and cross-linking studies only

52 estosterone were elevated in male and female GC-A(8E/8E) mice, but aldosterone was only increased in

53 FLAG-GC-A bound ANP identically with wild-type GC-A and was i

54 ANP did not increase internalization of FLAG-GC-A.

55 othelium-restricted deletion of GC-A (floxed GC-A x tie2-Cre: endothelial cell (EC) GC-A knockout (KO

56 (KO)); and (iii) control littermates (floxed GC-A mice with normal GC-A expression levels).

57 hether hsp90 plays a role as a chaperone for GC-A, the membrane guanylate cyclase that acts as a rece

58 lases (HDACs) in regulating Npr1 (coding for GC-A/NPRA) gene transcription, using primary mouse mesan

59 ble for regulating the Npr1 gene (coding for GC-A/NPRA) transcription are not well understood.

60 iuretic peptide (ANP), a proposed ligand for GC-A, has been suggested as critical for the maintenance

61 Heart weight to body weight ratios for GC-A(8E/8E) male, but not female, mice were 12% lower wi

62 ncreased internalization is not required for GC-A down-regulation.

63 This suggested that hsp90 was required for GC-A processing and/or stability.

64 increased in male but not female hearts from GC-A(8E/8E) mice.

65 increased in heart and kidney membranes from GC-A(8E/8E) mice.

66 h hypertension or heart failure, to generate GC-A-mediated cGMP ex vivo.

67 ocyte size was larger (approximately 20%) in GC-A null than in wild-type animals.

68 e, but the rapid increases were abolished in GC-A-deficient animals.

69 , but the effects of CNP were not altered in GC-A null mice.

70 ut 24 nM, but failed to relax such aortas in GC-A null mice, even at micromolar concentrations.

71 Although mice deficient in GC-A display an elevated blood pressure, the resultant c

72 uential deletion of individual glutamates in GC-A-8E progressively increased the Km Double Ala substi

73 Despite the marked reduction of GC-A mRNA in GC-A KO podocytes to 1% of the control level, Podo-GC-A

74 Km 23- to 70-fold but the same mutations in GC-A-8E only increased the Km 8-fold, consistent with on

75 anges in sodium excretion or urine output in GC-A-deficient mice.

76 iuresis in wild-type mice but no response in GC-A null animals.

77 ates for known phosphorylation sites to make GC-A(8E/8E) mice that express an enzyme that cannot be i

78 additional substitution for Ser-473 to make GC-A-8E resulted in the same Vmax, Km, and EC50 as the p

79 Adding more glutamates to make GC-A-9E or GC-A-10E had little effect on activity, and s

80 essure and heart rate were unchanged in male GC-A(8E/8E) mice.

81 lasma cGMP and decreased aldosterone in male GC-A(WT/WT) and GC-A(8E/8E) mice at 15 min, but only GC-

82 y and development of an oral, small molecule GC-A PAM that holds great potential as a therapeutic for

83 nd we successfully identified small molecule GC-A positive allosteric modulator (PAM) scaffolds.

84 ol littermates (floxed GC-A mice with normal GC-A expression levels).

85 eability effects of paracrine endothelial NP/GC-A/cGMP signaling and facilitate neutrophil extravasat

86 necrosis factor-alpha-induced endothelial NP/GC-A/cGMP/PDE2 signaling impairs endothelial barrier fun

87 However, NP/GC-A/cGMP signaling protects podocyte integrity under pa

88 retic peptide (ANP)-stimulated activation of GC-A.

89 of GC-B and 7-fold less potent activator of GC-A compared with wild type.

90 mice with endothelium-restricted deletion of GC-A (floxed GC-A x tie2-Cre: endothelial cell (EC) GC-A

91 with endothelial-restricted inactivation of GC-A (knockout mice with endothelial GC-A deletion) or c

92 Internalization of GC-A and NPR-C is poorly understood, in part, because pr

93 siRNA-mediated knockdown of GC-A, VASP, or PKG abolishes ANP-induced VASP Ser-239 ph

94 ly suppressed by siRNA-mediated knockdown of GC-A.

95 ed mice with a podocyte-specific knockout of GC-A (Podo-GC-A KO).

96 omain II or the aspartate in the DYG-loop of GC-A and GC-B failed to decrease enzyme phosphate conten

97 ent study was to delineate the mechanisms of GC-A/natriuretic peptide receptor-A (GC-A/NPRA) gene (Np

98 e significantly reduced by overexpression of GC-A, and this reduction was independent of genotype.

99 Thus, the phosphorylation of GC-A correlates with the acquisition of ligand sensitivi

100 ytic aspartate, are conserved in the PKDs of GC-A and GC-B.

101 Despite the marked reduction of GC-A mRNA in GC-A KO podocytes to 1% of the control leve

102 es have a daul function in the regulation of GC-A through both phosphorylation of and binding to regu

103 inactivates the enzyme, the significance of GC-A phosphorylation to heart structure and function rem

104 reduces GTP binding to the catalytic site of GC-A and GC-B and that ATP increases the magnitude of th

105 ent without reducing the maximal velocity of GC-A and GC-B.

106 tracks extracellular FLAG-tagged versions of GC-A and NPR-C independently of each other and ligand fo

107 WT) and GC-A(8E/8E) mice at 15 min, but only GC-A(8E/8E) mice had elevated levels of plasma cGMP and

108 Adding more glutamates to make GC-A-9E or GC-A-10E had little effect on activity, and sequential d

109 GC-A in the cardiac myocytes of wild-type or GC-A null animals.

110 nes from NIH 3T3 cells stably overexpressing GC-A were incubated with ATP, AMPPNP, or ATPgammaS, only

111 Here we overproduce GC-A in the cardiac myocytes of wild-type or GC-A null a

112 ALDO pod GC-A cKO mice demonstrated increased urinary albumin exc

113 ocytes, we generated podocyte-specific (pod) GC-A conditional KO (cKO) mice.

114 h a podocyte-specific knockout of GC-A (Podo-GC-A KO).

115 al hypertension of similar magnitude in Podo-GC-A KO mice and controls.

116 O podocytes to 1% of the control level, Podo-GC-A KO mice and control littermates did not differ in B

117 Concomitant treatment of Podo-GC-A KO mice with the TRPC channel blocker SKF96365 mark

118 -induced calcium influx in podocytes of Podo-GC-A KO mice.

119 However, only Podo-GC-A KO mice developed massive albuminuria (controls: 35

120 The particulate guanylyl cyclase A receptor (GC-A), via activation by its endogenous ligands atrial n

121 When cultured cells expressing recombinant GC-A were treated with geldanamycin, an inhibitor of hsp

122 blishment of an in vitro system to sensitize GC-A demonstrates that adenine nucleotides have a daul f

123 with ATP or ATPgammaS effectively sensitized GC-A to ligand stimulation over prolonged periods of tim

124 tive that thiophosphorylation had sensitized GC-A to ligand and adenine nucleotide binding.

125 s for all seven chemically identified sites (GC-A-7E) had a Km approximately 10-fold higher than phos

126 e-infused, and high salt-fed (ALDO) systemic GC-A KO mice with enhanced phosphorylation of p38 mitoge

127 educe that NPR-C is internalized faster than GC-A and that increased internalization is not required

128 peptide acted through a receptor other than GC-A.

129 The data suggest that GC-A is regulated by hsp90 complexes similar to those in

130 Introduction of the GC-A transgene did not alter blood pressure or heart rat

131 However, introduction of the GC-A transgene reduced cardiac myocyte size in both wild

132 orally bioavailable and directly targets the GC-A to potentiate cGMP has yet to be discovered.

133 ructures regulate blood pressure through the GC-A receptor.

134 Thus, the GC-A represents an important molecular therapeutic targe

135 natriuretic peptides (ANP, BNP) act through GC-A whereas another (CNP) acts through another receptor

136 from the heart function exclusively through GC-A.

137 ANP, therefore, appears to signal through GC-A in the kidney.

138 Thus, GC-A appears dispensable for regulation of sodium/water

139 d selectively enhances the binding of ANP to GC-A.

140 binding analysis confirmed MCUF-651 binds to GC-A and selectively enhances the binding of ANP to GC-A

141 studies ascribed NPR-C-mediated processes to GC-A.

142 sure concomitantly reduced surface and total GC-A levels, consistent with rapid exchange of extracell

143 AG-GC-A bound ANP identically with wild-type GC-A and was internalized slowly (0.5%/min), whereas FLA

144 re incubated with ATPgammaS and then washed, GC-A now became sensitive to ANP/AMPPNP stimulation, sug

145 lay a role in arteriopathies in women, while GC-A agonists may provide effective therapy for arteriti

146 The association of hsp90 and p50(cdc37) with GC-A was dependent on the kinase homology domain of this

147 ld higher than phosphorylated wild-type (WT) GC-A, but one additional substitution for Ser-473 to mak

148 and either Thr-500, Ser-510 or Thr-513 in WT-GC-A increased the Km 23- to 70-fold but the same mutati