ABCMdb

ABCC7 p.Ser813Ala

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Publications

PMID: 11053017 [PubMed] Baldursson O et al: "Contribution of R domain phosphoserines to the function of CFTR studied in Fischer rat thyroid epithelia."

No. Sentence Comment

56 In each of the serine to alanine mutants, S660A, S737A, S795A, and S813A, alanine replaced serine at the designated residue. Login to comment
107 The S660A and S813A variants generated small but significant cAMP-stimulated Cl- currents. Login to comment
112 The S660A and S813A variants generated currents, but they were no greater than those obtained with SQuad-A. Login to comment
186 Mutation of either residue alone significantly decreased current; with maximal stimulation by cAMP agonists, neither the S660A nor the S813A mutant gave more current than the S-Quad-A Fig. 5. Login to comment
195 With submaximal cAMP agonists, S660A- and S813A-generated current was also reduced, but it was slightly greater than that obtained with S-Quad-A. Login to comment

PMID: 15155835 [PubMed] Ai T et al: "Capsaicin potentiates wild-type and mutant cystic fibrosis transmembrane conductance regulator chloride-channel currents."

No. Sentence Comment

142 We further explored the action of capsaicin using CFTR mutants whose eight major PKA consensus serines are substituted with alanine (S660A, S686A, S700A, S712A, S737A, S768A, S795A, and S813A), so called S-oct-A or 8SA. Login to comment

PMID: 9922377 [PubMed] Gadsby DC et al: "Control of CFTR channel gating by phosphorylation and nucleotide hydrolysis."

No. Sentence Comment

183 Nevertheless,site Ser-813 seems to be effectively canceled by phosphorylation of the major inhibitory site Ser-768, since the dou- PKC stimulation or application has invariably been found to potentiate subsequent CFTR channel activation byble mutant S768A/S813A had a K0.5 for IBMX comparable to that of wild-type CFTR channels (220). Login to comment
386 A qualitatively similar slowing of the activation of macroscopic CFTR conductance, relative totural and functional considerations have led to the suggestion that the NBDs of ABC transporters might more closely wild-type CFTR, was observed for mutant S813A channels expressed in oocytes, but because those activation ratesresemble the catalytic sites of GTPases than of ion-motive ATPases (27, 68, 124). Login to comment

PMID: 19328185 [PubMed] Hegedus T et al: "Role of individual R domain phosphorylation sites in CFTR regulation by protein kinase A."

No. Sentence Comment

152 Membranes isolated from BHK cells expressing S700A (A), S737A (B), S768A (C), or S813A (D) were subjected to phosphorylation for 0, 2, 4, 8, 16, 32, and 64 min. Login to comment
151 Membranes isolated from BHK cells expressing S700A (A), S737A (B), S768A (C), or S813A (D) were subjected to phosphorylation for 0, 2, 4, 8, 16, 32, and 64 min. Login to comment

PMID: 9305845 [PubMed] Winter MC et al: "Stimulation of CFTR activity by its phosphorylated R domain."

No. Sentence Comment

185 Number of experiments for b/c and d were: 17/6 for wild-type (WT), 8 for wild-type (-PKA), 8/7 for S660A, 5/5 for S737A, 8/6 for S795A, 9/7 for S813A, and 11/4 for S-Quad-A. Login to comment
188 0.0 0.1 0.2 0.3 0.4 0.5 0.6 Po 0 0.2 0.4 0.6 0.8 1 10 [ATP] (mM) S813A S795A S737A S660A WT (-PKA) WT Figure 4 Effect of ATP concentration on Po of CFTR containing phosphorylation site mutations. Login to comment

PMID: 9252549 [PubMed] Wilkinson DJ et al: "CFTR activation: additive effects of stimulatory and inhibitory phosphorylation sites in the R domain."

No. Sentence Comment

87 S686A 0.75 ?I 0.07 9 N PKC site S700A 0.86 t 0.07* 12 S ++ +++ S712A 0.67 t 0.12 9 N - ++ S737A 0.35 5 0.05* 8 I +++ +++ S768A 0.09 IT 0.03* 8 I - ++++ S795A 1.24 + 0.22* 9 S +++ ++++ S813A 3.18-+0.36* 6 S ++++ ++ Values of half-maximal inhibition constant for activation (KA) are means + SE by 3-isobutyl-1-methylxanthine (IBMX) obtained for wild-type cystic fibrosis transmembrane conductance regulator (CFTR) and variants with single serine-to-alanine substitutions. Login to comment
107 Symbols show averaged IBMX dose-response data for wild-type CFTR and several single-site serine-to-alanine mutants (S660A, S737A, S768A, S795A, and S813A). Login to comment
119 100 i - wild type m m - S813A 0 S768,813A A S737,795,813A v S660,795,813A S660,737,795, E Fig. 2. Login to comment
125 For reference, the activation profiles of wild-type CFTR and single-site mutant S813A are shown by the thick continuous and dashed curves, respectively. Login to comment
129 One triple mutant (S737, 795, 813A) eliminated one inhibitory site (S737) and two stimulatory sites (S795 and S813), and the sensitivity to activation was not significantly different from that of the single-site mutant, S813A, as if the effects of eliminating S737 and S795 roughly canceled. Login to comment
130 In contrast, the other triple mutant (S660, 795, 813A), which lacked the three strongest stimulatory sites, exhibited a KA that was substantially greater than that of S813A, indicating that the effects produced by eliminating each of these sites summed such that the activation sensitivity of CFTR was considerably impaired. Login to comment
132 The value of KA was significantly reduced compared with that observed with the triple mutant (S660, 795,813A) but was significantly greater than that of S813A, again indicating a summation of effects such that the elimination of S737 partially ameliorated the reduced activation sensitivity produced by eliminating S660 and S795. Login to comment
136 26 7 S813A 533t75* 8 2.7 + 0.3* 173515* 9 Summary of activation and deactivation parameters (means 5 SE) for wild-type CFTR and the alanine substitution mutants of the strongest inhibitory (S768) and stimulatory (S813) phosphorylation sites. Login to comment
145 The data displayed in Table 3 indicate that the rate of approach to the activated steady state was decreased by -20% in the S813A construct, whereas the deactivation parameters, the latency, and the subsequent rate of exponential decline (*&E) indicated a destabilization of the active state. Login to comment
148 Deactivation parameters were consistent with stabilization of the active state; the latency was double that of wild-type CFTR, Y 80 zl 2 60 0 wt 0 S813A B 5 minutes z 80 iii n 60 0 10 20 30 40 50 minutes Fig. 3. Login to comment
149 Representative time courses for the activation (A) and deactivation (B) of wild-type (wt) CFTR and single-site mutants S768A and S813A after, respectively, exposure to 10 PM forskolin and 5 mM IBMX and the removal of these drugs from the perfusate. Login to comment

PMID: 7690753 [PubMed] Rich DP et al: "Regulation of the cystic fibrosis transmembrane conductance regulator Cl- channel by negative charge in the R domain."

No. Sentence Comment

48 Mutants are named by the number of the amino acid residue preceded by the wild-type amino acid and followed bythe aminoacid to which the residue was changed using the single-letter aminoacid code; for example,inthemutant CFTR S66OA,S737A,S795A,S813A (also called CFTR S-Quad-A for convenience), alanines are substituted for serines at amino acid positions 660, 737, 795, and 813. Login to comment
66 In vivo phosphorylation ofwild-typeand mutant CFTR CFTR S-Quad-A (S66OA,S737A,S795A,S813A), or CFTR S-Oct-A (S660A,S686A,S700A,S712A,S737A,S768A,S795A,S813A) weretran- siently expressed in HeLa cells. Login to comment
89 Functional assay of CFTR S-Quad-Aby SPQ fluorescence.The changein SPQfluorescenceis shown for HeLa cellsexpress- ing wild-type CFTR (n = 53, where n = number of cells) or CFTR S-Quad-A tS660A,S737A,S795A,S813A)(n = 53) and for virus only- infectedcells(no plasmid) as a negative control (n = 47). Login to comment
94 Single-channelanalysis of gle-channel traces from an excised, in-cell expressing CFTR S-Quad-A side-out membrane patch from a HeLa (S660A,S737A,S795A,S813A).Each trace is about 14s long. Login to comment
109 RESULTS Serine-to-Alanine Substitutionsin the R Domain Do NotAbolish CFTR Cl- Channel Activity-& a firststep to address the possibility that additional PKA phosphorylation sites might regulate CFTR, we asked whether the S-Quad-A (S660A,S737A,S795A,S813A) mutant could still bephosphorylated in vivofollowing CAMPstimulation. Login to comment
121 The single-channel open-stateprobabil- ity for wild-type CFTR (n = 14), CFTR S-Quad-A (S600A,S737A, S712A,S737A,S768A,S795A,S813A) (n = 7), or CFTR S-Oct-D (S660D,S686D,S700D,S712D,S737D,S768D,S795D,S813D)(n = 7in ATP alone (-PKA);n = 10 inPKA and ATP (+PkX))C1-channels was determined as described under "Experimental Procedures." Login to comment
123 S795A,S813A) (n = 5). Login to comment
139 CFTR (A) or CFTR S-Quad-A (S660A,S737A,S795A,S813A)( B )were transiently expressed inCOS-7 cells. Login to comment
176 COS-7 cells were mock- (S660A,S686A,S700A,S712A,S737A,S768A,S795A,S813A)(C). Login to comment
47 Mutants are named by the number of the amino acid residue preceded by the wild-type amino acid and followed bythe aminoacid to which the residue was changed using the single-letter amino acid code; for example,inthemutant CFTR S66OA,S737A,S795A,S813A (also called CFTR S-Quad-A for convenience), alanines are substituted for serines at amino acid positions 660, 737, 795, and 813. Login to comment
65 In vivo phosphorylation ofwild-typeand mutant CFTR CFTR S-Quad-A (S66OA,S737A,S795A,S813A), or CFTR S-Oct-A (S660A,S686A,S700A,S712A,S737A,S768A,S795A,S813A) were transiently expressed in HeLa cells. Login to comment
95 Single-channelanalysis of gle-channel traces from an excised, in-cell expressing CFTR S-Quad-A side-out membrane patch from a HeLa (S660A,S737A,S795A,S813A).Each trace is about 14s long. Login to comment
111 RESULTS Serine-to-Alanine Substitutions in the R Domain Do NotAbolish CFTR Cl-Channel Activity-& a firststep to address the possibility that additional PKA phosphorylation sites might regulate CFTR, we asked whether the S-Quad-A (S660A,S737A,S795A,S813A) mutant could still be phosphorylated in vivofollowing CAMPstimulation. Login to comment
125 S795A,S813A) (n = 5). Login to comment
142 CFTR (A) or CFTR S-Quad-A (S660A,S737A,S795A,S813A) ( B )were transiently expressed inCOS-7 cells. Login to comment
180 COS-7 cells were mock- (S660A,S686A,S700A,S712A,S737A,S768A,S795A,S813A) (C). Login to comment

PMID: 7684377 [PubMed] Chang XB et al: "Protein kinase A (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites."

No. Sentence Comment

37 The following mutations were introduced into CFTR, S422A (TCT to GCT), S660A (TCA to GCA), S686A (TCT to GCT), S700A (TCT toGCT), S712A (TCC to GCC), S737A (TCC to GCC), S768A (TCT toGCT), T788A (ACAto GCA),S795A (TCA to GCA), S813A (TCA to GCA), S660E (TCA to GAA),S737E (TCC to GAG), S795E (TCA to GAA), and S813E (TCA to GAA). Login to comment
39 The counterparts of CFTR cDNA in pUCF2.5 were replaced by PCR- mutated versions by interchange of the following fragments, S660A, DraIIIIEcoRI fragment, S737A, EcoRIIHpaIfragment, S795A and S813A,StyIIStyI fragment(Fig. 1A). Login to comment

PMID: 23760269 [PubMed] Billet A et al: "Role of tyrosine phosphorylation in the muscarinic activation of the cystic fibrosis transmembrane conductance regulator (CFTR)."

No. Sentence Comment

102 Carbachol Stimulates CFTR through PKA and Non-PKA Signaling Pathways-To explore PKA-independent regulation of CFTR without using inhibitors that might have confounding effects on other pathways, we studied the activation of 15SA-CFTR (S422A/S660A/S670A/S686A/T690A/S700A/S712A/ S737A/S753A/S768A/T787A/T788A/S790A/S795A/S813A). Login to comment