ABCC7 p.Thr665Ala
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PMID: 11331356
[PubMed]
Button B et al: "PKC-mediated stimulation of amphibian CFTR depends on a single phosphorylation consensus site. insertion of this site confers PKC sensitivity to human CFTR."
No.
Sentence
Comment
70
The following primers were used (only sense primer is shown, with mutations underlined): 5Ј-TAATAACTGAGGCCCTGAGACGATGCT-3Ј (Thr665 to Ala, T A B L E I Conductances of Xenopus Oocytes Expressing CFTR Unstimulated cAMP-stimulated 24 h 48 h 72 h Mock 2.9 Ϯ 1.7 (11) 2.7 Ϯ 1.4 (4) 2.9 Ϯ 1.6 (4) 3.2 Ϯ 2.1 (3) hCFTR 4.2 Ϯ 3.8 (35) 426 Ϯ 60 (16) 685 Ϯ 131 (13) 1,147 Ϯ 217 (6) XCFTR 3.1 Ϯ 2.1 (34) 214 Ϯ 50 (14) 1,009 Ϯ 268 (14) 2,028 Ϯ 217 (6) Xenopus laevis oocytes were injected with water (mock), hCFTR cRNA, or XCFTR cRNA, and their membrane conductances were measured at 24, 48, or 72 h after injection with and without stimulation with cAMP cocktail.
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ABCC7 p.Thr665Ala 11331356:70:136
status: NEW207 Thr665 Is Essential for the Response to PKC Activation of XCFTR To investigate the roles of the unique PKC consensus sites (Thr665 and Ser694), we expressed the mutant lacking both sites (T665A/S694A-XCFTR) in Xenopus oocytes.
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ABCC7 p.Thr665Ala 11331356:207:188
status: NEW215 (B) I-V plot from a cell expressing the double knockout of unique PKC consensus phosphorylation sites (T665A/S694A-XCFTR).
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ABCC7 p.Thr665Ala 11331356:215:103
status: NEW221 In contrast, in oocytes expressing T665A-XCFTR, PMA failed to produce full current activation (Fig. 8, B and C).
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ABCC7 p.Thr665Ala 11331356:221:35
status: NEW223 Thus, it is possible that the reduced activation by PMA of XCFTR mutants containing the Thr665 to Ala mutation is not absolute, but relative to the activation by PKA stimulation (i.e., the mutation increases the activation of CFTR by cAMP).
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ABCC7 p.Thr665Ala 11331356:223:88
status: NEW224 The data in Fig. 9 indicate that this is not the case because the cAMP-activated currents are not different among oocytes expressing wild-type XCFTR, S686A/ S790A-XCFTR, or T665A/S694A-XCFTR.
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ABCC7 p.Thr665Ala 11331356:224:173
status: NEW245 The mutation Thr665 to Ala does not increase cAMP-activated XCFTR currents.
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ABCC7 p.Thr665Ala 11331356:245:13
status: NEW246 Time course of the currents after stimulation with the cAMP cocktail in oocytes injected with wild-type XCFTR, S686A/S790A-XCFTR, or T665A/S694A-XCFTR cRNAs.
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ABCC7 p.Thr665Ala 11331356:246:133
status: NEW248 The data show that the Thr665 to Ala mutation has no significant effect on the level of cAMP-activated currents.
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ABCC7 p.Thr665Ala 11331356:248:23
status: NEW
PMID: 15229107
[PubMed]
Chen Y et al: "Mechanism of activation of Xenopus CFTR by stimulation of PKC."
No.
Sentence
Comment
211
A nonspecific stimulatory effect of high PMA concentration can be ruled out because the same concentration of 4␣-PMA, an inactive form of PMA, had no effect, and the mutation T665A reduced the activation of XCFTR by PMA by ϳ75% (6).
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ABCC7 p.Thr665Ala 15229107:211:182
status: NEW
PMID: 15282191
[PubMed]
Chen Y et al: "Potentiation of effect of PKA stimulation of Xenopus CFTR by activation of PKC: role of NBD2."
No.
Sentence
Comment
207
We found that the activation by PKC stimulation was abolished in the mutant T665A XCFTR and transferred to the hCFTR mutant TLHR 3 TLRR (7).
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ABCC7 p.Thr665Ala 15282191:207:76
status: NEW