ABCC7 p.His954Cys

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PMID: 20952391 [PubMed] Wang G et al: "State-dependent regulation of cystic fibrosis transmembrane conductance regulator (CFTR) gating by a high affinity Fe3+ bridge between the regulatory domain and cytoplasmic loop 3."
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155 Similarly, disulfide bond cross-linking of H950C or H954C to S832C, H775C, or D836C also inhibited channel activity, whereas single cysteine mutants were not affected by diamide (Fig. 5F and supplemental Fig. S1).
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ABCC7 p.His954Cys 20952391:155:52
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156 However, both diamide and DTT failed to exert any effect on a H954C/D828C mutant possibly because of a poor orientation or a long distance between these two engineered cysteines (Fig. 5F and supplemental Fig. S1).
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ABCC7 p.His954Cys 20952391:156:62
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157 Additionally, small inhibition of the current by diamide (Fig. 5F) was not due to a background noise induced by a low current amplitude in the Cys-free constructs because a WT CFTR-based H954C mutant, which contains Cys-832 in the R domain, also exhibited a clear inhibition of the current by diamide (Fig. 5D).
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ABCC7 p.His954Cys 20952391:157:187
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227 A-E, macroscopic currents across inside-out membrane patches excised from transfected HEK-293T cells expressing mutants H950C/S832C/V510A (A), H950C/V510A (B), S832C/V510A (C), H954C (D), and the WT hCFTR construct (E).
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ABCC7 p.His954Cys 20952391:227:177
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PMID: 21059651 [PubMed] Wang G et al: "The inhibition mechanism of non-phosphorylated Ser768 in the regulatory domain of cystic fibrosis transmembrane conductance regulator."
No. Sentence Comment
118 Supporting this argument, internal diamide also inhibited activity of a mutant S768C/K951C, S768C/H954C, or S768C/ S955C, and inhibition was reversed by 4-6 mM DTT (Fig. 2E).
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ABCC7 p.His954Cys 21059651:118:98
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119 In sharp contrast, both diamide and DTT had no effect on such CFTR constructs as K951C, H954C, and S955C.
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ABCC7 p.His954Cys 21059651:119:88
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122 Similarly, diamide also suppressed channel activity of mutants S737C/H954C, S737C/S955C, and S737C/Q958C, and suppression was reversed by DTT (Fig. 2E).
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ABCC7 p.His954Cys 21059651:122:69
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125 Fig. 3 demonstrates that a CFTR construct with a single cysteine S768C, S737C, H950C, or H954C exhibited a clear single band no matter whether diamide or DTT was added.
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ABCC7 p.His954Cys 21059651:125:89
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126 In sharp contrast, CFTR constructs with a cysteine pair (Cys-free background), S737C/H950C, S737C/H954C, S768C/H950C, and S768C/H954C, exhibited an additional cross-linked (X-linked) band because it was induced by diamide but was weakened by DTT.
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ABCC7 p.His954Cys 21059651:126:98
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ABCC7 p.His954Cys 21059651:126:128
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127 In contrast, the H950C/V956C mutant exhibited no X-linked band possibly because of a poor relative orientation between H954C and V956C.
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ABCC7 p.His954Cys 21059651:127:119
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128 Therefore, a disulfide bond can be formed between H950C (or H954C) and S768C or between H954C (or H950C) and S737C.
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ABCC7 p.His954Cys 21059651:128:60
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ABCC7 p.His954Cys 21059651:128:88
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143 In contrast, curcumin had no such effect on S737A and H954A mutants, suggesting that they may be weak inhibitory residues, although disulfide cross-linking of S737C to H954C strongly inhibited channel activity (Figs. 2E and 4E).
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ABCC7 p.His954Cys 21059651:143:168
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