ABCC7 p.Asn268Cys
| Predicted by SNAP2: | A: D (63%), C: D (75%), D: D (59%), E: D (75%), F: D (91%), G: N (72%), H: D (63%), I: D (91%), K: D (80%), L: D (91%), M: D (85%), P: D (75%), Q: D (66%), R: D (80%), S: N (53%), T: D (59%), V: D (85%), W: D (91%), Y: D (80%), |
| Predicted by PROVEAN: | A: D, C: D, D: D, E: D, F: D, G: N, H: D, I: D, K: D, L: D, M: D, P: D, Q: D, R: D, S: N, T: D, V: D, W: D, Y: D, |
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[hide] Multiple membrane-cytoplasmic domain contacts in t... J Biol Chem. 2008 Sep 26;283(39):26383-90. Epub 2008 Jul 25. He L, Aleksandrov AA, Serohijos AW, Hegedus T, Aleksandrov LA, Cui L, Dokholyan NV, Riordan JR
Multiple membrane-cytoplasmic domain contacts in the cystic fibrosis transmembrane conductance regulator (CFTR) mediate regulation of channel gating.
J Biol Chem. 2008 Sep 26;283(39):26383-90. Epub 2008 Jul 25., 2008-09-26 [PMID:18658148]
Abstract [show]
The cystic fibrosis transmembrane conductance regulator (CFTR) is a unique ATP-binding cassette (ABC) ion channel mutated in patients with cystic fibrosis. The most common mutation, deletion of phenylalanine 508 (DeltaF508) and many other disease-associated mutations occur in the nucleotide binding domains (NBD) and the cytoplasmic loops (CL) of the membrane-spanning domains (MSD). A recently constructed computational model of the CFTR three-dimensional structure, supported by experimental data (Serohijos, A. W., Hegedus, T., Aleksandrov, A. A., He, L., Cui, L., Dokholyan, N. V., and Riordan, J. R. (2008) Proc. Natl. Acad. Sci. U. S. A. 105, 3256-3261) revealed that several of these mutations including DeltaF508 disrupted interfaces between these domains. Here we have used cysteine cross-linking experiments to verify all NBD/CL interfaces predicted by the structural model and observed that their cross-linking has a variety of different effects on channel gating. The interdomain contacts comprise aromatic clusters important for stabilization of the interfaces and also involve the Q-loops and X-loops that are in close proximity to the ATP binding sites. Cross-linking of all domain-swapping contacts between NBDs and MSD cytoplasmic loops in opposite halves of the protein rapidly and reversibly arrest single channel gating while those in the same halves have lesser impact. These results reinforce the idea that mediation of regulatory signals between cytoplasmic- and membrane-integrated domains of the CFTR channel apparently relies on an array of precise but highly dynamic interdomain structural joints.
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No. Sentence Comment
84 The successful cross-linking of the residue pairs N268C (CL2) and F1294C (NBD2) by the MTS reagents of various spacer arm lengths confirmed this contact (Fig. 2A).
X
ABCC7 p.Asn268Cys 18658148:84:50
status: NEW90 Cys pair cross-linking experiments showed that indeed E543C could be cross-linked with both T966C (CL3) and T1057C (CL4, Fig. 2B), while D1341C was in close contact with both L172C (CL1) and N268C (CL2, Fig. 2C).
X
ABCC7 p.Asn268Cys 18658148:90:191
status: NEW100 A, 276C/Y1307C and N268C/F1292C at the CL2/NBD2 interface involving an aromatic cluster and the Q-loop.
X
ABCC7 p.Asn268Cys 18658148:100:19
status: NEW102 C, L172C/D1341C at the CL1/NBD2 interface and N268C/D1341C at the CL2/NBD2 interface at the X-loop of NBD2.
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ABCC7 p.Asn268Cys 18658148:102:46
status: NEW150 As shown in Fig. 5, treatment of the cells with PKA stimulation mixture before and during cross-linking had no effect on cross-linking between either M498C/G1061C (Fig. 5A, left panel) or N268C/F1294C (Fig. 5B, left panel), although phosphorylation-sensitive CFTR antibody mAb 450 confirmed the phosphorylation of CFTR (Fig. 5, A and B, right panels).
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ABCC7 p.Asn268Cys 18658148:150:188
status: NEW166 Cys pairs were introduced at: A interface between NBD1 Q-loop and CL4 (M498C/G1061C); B, interface between NBD2 Q-loop and CL2 (N268C/ F1294C); or C, both Q-loops (W496C and K1292C).
X
ABCC7 p.Asn268Cys 18658148:166:128
status: NEW