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- W2088647578 abstract "Most ATP-binding cassette (ABC) transporters have two intracellular nucleotide-binding domains (NBDs) and two membrane-spanning domains (MSDs). Each MSD consists of six helical segments probably extended to four gating-controled cytoplasmic loops (CLs). However, the cyctic fibrosis transmembrane conductance regulator (CFTR) has an additional regulatory (R) domain (645-838). It has fourteen PKA phosphorylation sites exerting stimulatory or inhibitory effects on channel activity. My previous studies suggested that S768, a well-known inhibitory PKA phosphorylation site, may prevent channel opening by a putative Fe3+ bridge or an H-bond between the R domain and CL3. Here an electrostatic attraction between the R domain and CL1 or CL3 was reported as a new approach to regulating channel gating dually. On the one hand, diamide-induced disulfide bond crosslinking of K946C to D836C, confirmed by SDS-PAGE mobility, inhibited channel opening, and inhibition was reversed by DTT. However, both K946A and D835A or D836A clearly increased the sensitivity of channel activity to PKA phosphorylation while K951A failed to. Thus, an electrostatic attraction between K946 and D835 or D836 may stop channel opening primarily by preventing an interaction of CL3 with CL1. On the other hand, diamide-induced disulfide bond crosslinking of S700C or S795C to K162C or K163C, which was confirmed by SDS-PAGE mobility upon pretreatment of CFTR-expressing HEK-293T cells with forskolin, increased a channel open probability, and the effect was reversed by DTT. However, S700A, S795A and K162A enhanced PKA dependence of channel activity while K163A did not. Therefore, an electrostatic attraction between K162 and phosphorylated S700 or S795 may promote channel opening possibly by stabilizing the interaction between CL1 and CL3." @default.
- W2088647578 created "2016-06-24" @default.
- W2088647578 creator A5088423634 @default.
- W2088647578 date "2011-02-01" @default.
- W2088647578 modified "2023-09-26" @default.
- W2088647578 title "Dual Regulation of Intracellular CFTR Gating by the R Domain" @default.
- W2088647578 doi "https://doi.org/10.1016/j.bpj.2010.12.1661" @default.
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