ABCMdb

ABCC7 p.His950Cys

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Publications

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."

No. Sentence Comment

147 Fig. 5A shows that internal diamide (10 ␮M) suppressed ϳ30% of channel activity of a H950C/S832C/ V510A construct, and suppression was partially reversed by FIGURE 2. Login to comment
154 These observations suggest that disulfide bond cross-linking between H950C and S832C should inhibit channel activity. Login to comment
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). Login to comment
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). Login to comment

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

95 S768C/H950C was a representative example (Fig. 2B). Login to comment
99 B-D, macroscopic currents across inside-out membrane patches excised from transfected HEK-293T cells expressing mutants H950C/S768C (B), H950C (C), and S768C (D) by using a ramp protocol (Ϯ80 mV). Login to comment
108 F, unitary currents from a H950C/S768C construct in the presence of 20 ␮M diamide (b) and 4 mM DTT (c). Login to comment
115 In contrast, both diamide and DTT had no effect on H950C and S768C CFTR constructs (Fig. 2, C and D). Login to comment
116 These observations clearly suggest that a disulfide bond may be formed between S768C and H950C. Login to comment
120 Furthermore, channel activity of V769C/H950C was also inhibited by diamide (Fig. 2E), suggesting that phosphorylation of Ser768 may not affect the CL3-R interface. Login to comment
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. Login to comment
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. Login to comment
127 In contrast, the H950C/V956C mutant exhibited no X-linked band possibly because of a poor relative orientation between H954C and V956C. Login to comment
128 Therefore, a disulfide bond can be formed between H950C (or H954C) and S768C or between H954C (or H950C) and S737C. Login to comment
129 In order to address if the disulfide bond changes the gating kinetics, a two-channel recording of the H950C/S768C construct was done. Login to comment
162 Because disulfide cross-linking of S768C to H950C inhibited more channel activity than that of S768C to S955C (Fig. 2E), it is more possible for His950 to form an inhibitory H-bond with Ser768 . Login to comment
244 Although thiol-specific disulfide cross-linking of S768C to H950C or nearby cysteines inserted in CL3 inhibited channel activity primarily by stopping the channel from opening, an electrostatic expulsion between S768R/D and H950R/D clearly promoted channel opening even in the absence of ATP. Login to comment
288 Second, diamide-induced disulfide bond cross-linking of S768C to H950C or its neighboring cysteines, which was confirmed by the SDS-PAGE mobility (Fig. 3), inhibited channel activity (Fig. 2). Login to comment