ABCC7 p.Arg766Ala
| ClinVar: |
c.2297G>T
,
p.Arg766Met
?
, not provided
|
| CF databases: |
c.2297G>T
,
p.Arg766Met
(CFTR1)
D
, R766M was identified by non-radioactive SSCP and direct sequencing; it creates a FokI restriction site.
|
| Predicted by SNAP2: | A: D (85%), C: D (91%), D: D (95%), E: D (91%), F: D (91%), G: D (91%), H: D (91%), I: D (91%), K: D (75%), L: D (91%), M: D (66%), N: D (91%), P: D (91%), Q: D (85%), S: D (85%), T: D (85%), V: D (85%), W: D (95%), Y: D (91%), |
| Predicted by PROVEAN: | A: N, C: D, D: D, E: N, F: D, G: D, H: N, I: D, K: N, L: D, M: D, N: N, P: D, Q: N, S: N, T: D, V: D, W: D, Y: D, |
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[hide] Regulation of Activation and Processing of the Cys... J Biol Chem. 2012 Oct 11. Wang G, Duan DD
Regulation of Activation and Processing of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) by a Complex Electrostatic Interaction between the Regulatory Domain and Cytoplasmic Loop 3.
J Biol Chem. 2012 Oct 11., [PMID:23060444]
Abstract [show]
NEG2, a short C-terminal segment (817-838) of the unique regulatory (R) domain of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel, has been reported to regulate CFTR gating in response to cAMP-dependent R domain phosphorylation. The underlying mechanism, however, is unclear. Here, K946 of cytoplasmic loop 3 (CL3) is proposed as counter-ion of D835, D836 or E838 of NEG2 to prevent channel activation by PKA. R764 or R766 of the S768 phosphorylation site of the R domain is proposed to promote channel activation possibly by weakening the putative CL3-NEG2 electrostatic attraction. First, not only D835A, D836A and E838A but also K946A reduced the PKA dependent CFTR activation. Second, both K946D and D835R/D836R/E838R mutants were activated by ATP and curcumin to a different extent. Third, R764A and R766A mutants enhanced the PKA-dependent activation. On the other hand, it is very exciting that D835R/D836R/E838R and K946D/H950D and H950R exhibited normal channel processing and activity while D835R/D836R/E838R/K946D/H950D was misprocessed and silent in response to forskolin. Further, D836R and E838R played a critical role in the asymmetric electrostatic regulation of CFTR processing and S768 phosphorylation may not be involved. Thus, a complex interfacial interaction among CL3, NEG2 and the S768 phosphorylation site may be responsible for the asymmetric electrostatic regulation of CFTR activation and processing.
Comments [show]
None has been submitted yet.
No. Sentence Comment
11 First, not only D835A, D836A and E838A but also K946A reduced the PKA dependent CFTR activation.
X
ABCC7 p.Arg766Ala 23060444:11:17
status: NEW13 Third, R764A and R766A mutants enhanced the PKA-dependent activation.
X
ABCC7 p.Arg766Ala 23060444:13:17
status: NEW116 The effects of R764A and R766A on the PKA-dependent channel activation - Because both S768 and D836 are close to H950 of CL3, it is fitting to ask if R764, R765 and R766 form a salt bridge with the N-terminal of NEG2 to cause the asymmetry of the electrostatic regulation.
X
ABCC7 p.Arg766Ala 23060444:116:25
status: NEW118 Fig. 4D demonstrates that both R764A and R766A exhibited a normal channel density.
X
ABCC7 p.Arg766Ala 23060444:118:41
status: NEW121 The K1/2 for PKA activation of R764A and R766A increased from 10 units/ml to 25 and 44 units/ml, respectively.
X
ABCC7 p.Arg766Ala 23060444:121:41
status: NEW140 Finally, it is very exciting that the putative electrostatic attraction between K946D/H950D and D835R/D836R/E838R dramatically suppressed the channel activity by stopping the channel processing or opening(10).
X
ABCC7 p.Arg766Ala 23060444:140:25
status: NEW145 Our finding shows that R764A and R766A suppressed not the channel processing and the current density but the channel activation by PKA (Fig. 4-6).
X
ABCC7 p.Arg766Ala 23060444:145:33
status: NEW142 Fig. 4D demonstrates that both R764A and R766A exhibited a normal channel density.
X
ABCC7 p.Arg766Ala 23060444:142:41
status: NEW151 The currents mediated by S768D/K946D/H950D/D836R and S768D/K946D/H950D/E838R were statistically smaller than the S768D/K946D/H950D current (n afd; 3,*, p b0d; 0.05, unpaired t test); error bars, S.E. TABLE 1 WholecellcurrentsIm (picoamperes)andcapacitancesCm (picofarads) of HEK-293T cells expressing CFTR constructs in response to forskolin Constructs Stimulated Im Control Cm Stimulated Cm n WT-CFTR 701.6 afe; 9.0 17.4 afe; 0.9 16.4 afe; 1.1 3 D835R/D836R/E838R 539.5 afe; 5.3 15.0 afe; 1.0 16.2 afe; 0.8 3 Asymmetric Electrostatic Regulation of CFTR 40488 JOURNAL OF BIOLOGICAL CHEMISTRY VOLUME 287ߦNUMBER 48ߦNOVEMBER 23, 2012 at SEMMELWEIS UNIV OF MEDICINE on December , The K1/2 for PKA activation of R764A and R766A increased from 10 units/ml to 25 and 44 units/ml, respectively.
X
ABCC7 p.Arg766Ala 23060444:151:754
status: NEW173 The PKA-dependent activity of CFTR mutants R764A and R766A.
X
ABCC7 p.Arg766Ala 23060444:173:53
status: NEW174 A, unit channel currents across inside-out membrane patches excised from transfected HEK-293T cells expressing R766A by using a holding potential (af9;60 mV).
X
ABCC7 p.Arg766Ala 23060444:174:111
status: NEW179 B, PKA titration curves for CFTR mutants R764A and R766A (n afd; 3-5).
X
ABCC7 p.Arg766Ala 23060444:179:51
status: NEW184 Our finding shows that R764A and R766A suppressed not the channel processing and the current density but the channel activation by PKA (Figs. 4-6).
X
ABCC7 p.Arg766Ala 23060444:184:33
status: NEW