ABCC7 p.Ser422Asp
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No.
Sentence
Comment
141
water CFTR CFTR PIKfyve CFTR S318APIKfyve ΔF508CFTR PIKfyve C FTR +H 2 O C FTR +H 2 O C FTR +PIKfyve C FTR + S318A PIKfyve C FTR +PIKfyve C FTR + S422D SG K1 ~170 KDa ~170 KDa A B Fig. 4.
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ABCC7 p.Ser422Asp 19852935:141:152
status: NEW
No.
Sentence
Comment
44
Cell culture, lysis, protein solubilization and Western blotting The cell culture methods to create the stable CFTR-expressing cell lines (WT, F, WT S422A, WT S422D, WT S511A, WT S511D, WT T1471A and WT T1471D) and their culture protocols have been described recently [25].
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ABCC7 p.Ser422Asp 23067305:44:160
status: NEW191 Compared with each other S422D- and S422A-CFTR revealed no gross differences in fragmentation (Figures 7A-7D, compare lanes 3 and 4).
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ABCC7 p.Ser422Asp 23067305:191:25
status: NEW
PMID: 25384981
[PubMed]
Caohuy H et al: "Activation of 3-phosphoinositide-dependent kinase 1 (PDK1) and serum- and glucocorticoid-induced protein kinase 1 (SGK1) by short-chain sphingolipid C4-ceramide rescues the trafficking defect of DeltaF508-cystic fibrosis transmembrane conductance regulator (DeltaF508-CFTR)."
No.
Sentence
Comment
82
In Vitro Phosphorylation of CFTR-Wild-type and èc;F508-CFTR immunoprecipitates from untreated repaired and CFPAC-1 cell lysates, respectively, were washed with phosphorylation buffer (50 mM Tris, pH 7.5, 0.1 mM EGTA, 0.1% 2-mercaptoethanol) and then resuspended in 30 òe;l of the same buffer containing 100 ng of purified recombinant human S422D-SGK1 mutant protein (Millipore) and 1afb; Mg2af9; -ATP mixture (Millipore).
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ABCC7 p.Ser422Asp 25384981:82:348
status: NEW336 The in vitro kinase reaction mixture consists of active PDK1, mutant S422D-SGK1, and substrate CFTR that was immunoprecipitated from either untreated CFPAC-1 or repaired CFPAC-1 pLJ6 cells.
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ABCC7 p.Ser422Asp 25384981:336:69
status: NEW415 Phosphorylation of immunoprecipitated CFTR or phosphorylation at Thr256 of S422D-SGK1 in the whole reaction mixture(WRM)wasdetectedbyWesternblotting.D,effectofC4-CERonCFTRubiquitination.ThecelllysatespreparedasdescribedinBweresubjectedtoCFTR immunoprecipitation, followed by Western blot analysis for total CFTR expression or CFTR ubiquitination.
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ABCC7 p.Ser422Asp 25384981:415:75
status: NEW425 Furthermore, the importance of active SGK kinase for the addition of CFTR to the plasma membrane in Xenopus oocytes (50) is consistent with our observation that PDK1-activated S422D-SGK1 phosphorylates both wild-type and mutant CFTR.
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ABCC7 p.Ser422Asp 25384981:425:176
status: NEW