ABCC7 p.Arg1403Ala
| CF databases: |
c.4207A>G
,
p.Arg1403Gly
(CFTR1)
?
, The mutation on the other chromosome is unknown
|
| Predicted by SNAP2: | A: D (91%), C: D (91%), D: D (95%), E: D (95%), F: D (95%), G: D (95%), H: D (85%), I: D (95%), K: D (71%), L: D (91%), M: D (91%), N: D (91%), P: D (95%), Q: D (85%), S: D (91%), T: D (91%), V: D (95%), W: D (95%), Y: D (95%), |
| Predicted by PROVEAN: | A: D, C: D, D: D, E: D, F: D, G: D, H: D, I: D, K: N, L: D, M: D, N: D, P: D, Q: D, S: D, T: D, V: D, W: D, Y: D, |
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[hide] Localization of sequences within the C-terminal do... J Biol Chem. 2001 Jan 12;276(2):1291-8. Gentzsch M, Riordan JR
Localization of sequences within the C-terminal domain of the cystic fibrosis transmembrane conductance regulator which impact maturation and stability.
J Biol Chem. 2001 Jan 12;276(2):1291-8., 2001-01-12 [PMID:11022033]
Abstract [show]
Some disease-associated truncations within the 100-residue domain C-terminal of the second nucleotide-binding domain destabilize the mature protein (Haardt, M., Benharouga, M., Lechardeur, D., Kartner, N., and Lukacs, G. L. (1999) J. Biol. Chem. 274, 21873-21877). We now have identified three short oligopeptide regions in the C-terminal domain which impact cystic fibrosis transmembrane conductance regulator (CFTR) maturation and stability in different ways. A highly conserved hydrophobic patch (region I) formed by residues 1413-1416 (FLVI) was found to be crucial for the stability of the mature protein. Nascent chain stability was severely decreased by shortening the protein by 81 amino acids (1400X). This accelerated degradation was sensitive to proteasome inhibitors but not influenced by brefeldin A, indicating that it occurred at the endoplasmic reticulum. The five residues at positions 1400 to 1404 (region II) normally maintain nascent CFTR stability in a positional rather than a sequence-specific manner. A third modulating region (III) constituted by residues 1390 to 1394 destabilizes the protein. Hence the nascent form regains stability on further truncation back to residues 1390 or 1380, permitting some degree of maturation and a low level of cyclic AMP-stimulated chloride channel activity at the cell surface. Thus while not absolutely essential, the C-terminal domain strongly modulates the biogenesis and maturation of CFTR.
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No. Sentence Comment
40 F1413A/ L1414A/V1415A/I1416A/E1417A, 5Ј-GCTGGAATGCCAACAAGCTGC- GGCCGCAGCAGAGAACAAAGTGCGG-3Ј and 5Ј-CCGCACTTTGTTC- TCTGCTGCGGCCGCAGCTTGTTGGCATTCCAGC-3Ј; F1413A- /L1414A/V1415A/I1416A, 5Ј-GCTGGAATGCCAACAAGCTGCGGCCG- CAGAAGAGAACAAAGTGCGG-3Ј and 5Ј-CCGCACTTTGTTCTCTTC- TGCGGCCGCAGCTTGTTGGCATTCCAGC-3Ј; Q1411A/Q1412A, 5Ј-G- GATAGAAGCAATGCTGGAATGCGCAGCATTTTTGGTCATAGAAG-3 and 5Ј-CTTCTATGACCAAAAATGCTGCGCATTCCAGCATTGCTTCT- ATCC-3; F1413A/L1414A, 5Ј-GCTGGAATGCCAACAAGCTGCGGTCA- TAGAAGAGAACAAAGTGCG-3Ј and 5Ј-CGCACTTTGTTCTCTTCTA- TGACCGCAGCTTGTTGGCATTCCAGC-3Ј; L1414A/V1415A, 5Ј-GCT- GGAATGCCAACAATTTGCGGCCATAGAAGAGAACAAAGTGCGG-3Ј and 5Ј-CCGCACTTTGTTCTCTTCTATGGCCGCAAATTGTTGGCATT- CCAGC-3Ј; V1415A/I1416A, 5Ј-GGAATGCCAACAATTTTTGGCCGCA- GAAGAGAACAAAGTGCGGCAG-3Ј and 5Ј-CTGCCGCACTTTGTTCT- CTTCTGCGGCCAAAAATTGTTGGCATTCC-3Ј; E1417A/E1418A, 5Ј- GCCAACAATTTTTGGTCATAGCAGCGAACAAAGTGCGGCAGTAC- G-3Ј and 5Ј-CGTACTGCCGCACTTTGTTCGCTGCTATGACCAAAAA- TTGTTGGC-3Ј; F1413A, 5Ј-GCAATGCTGGAATGCCAACAAGCTTTG- GTCATAGAAGAGAAC-3Ј and 5Ј-GTTCTCTTCTATGACCAAAGCTT- GTTGGCATTCCAGCATTGC-3Ј; L1414A, 5Ј-GCTGGAATGCCAACAA- TTTGCGGTCATAGAAGAGAACAAAGTGCG-3Ј and 5Ј-CGCACTTTG- TTCTCTTCTATGACCGCAAATTGTTGGCATTCCAGC-3Ј; V1415A, 5Ј- GGAATGCCAACAATTTTTGGCCATAGAAGAGAACAAAGTGCGGC- AG-3Ј and 5Ј-CTGCCGCACTTTGTTCTCTTCTATGGCCAAAAATTGT- TGGCATTCC-3Ј; I1416A, 5Ј-GGAATGCCAACAATTTTTGGTCGCAG- AAGAGAACAAAGTGCGGCAG-3Ј and 5Ј-CTGCCGCACTTTGTTCTC- TTCTGCGACCAAAAATTGTTGGCATTCC-3Ј; E1417A, 5Ј-GCCAACA- ATTTTTGGTCATAGCAGAGAACAAAGTGCGGCAGTACG-3Ј and 5Ј- CGTACTGCCGCACTTTGTTCTCTGCTATGACCAAAAATTGTTGGC- 3Ј; C1400A/E1401A/H1402A/R1403A/I1404A, 5Ј-GCACAGTAATTCTC- GCTGCAGCCGCGGCAGAAGCAATGCTGGAATGCC-3Ј and 5Ј-GGC- ATTCCAGCATTGCTTCTGCCGCGGCTGCAGCGAGAATTACTGTG- C-3Ј; ⌬1400-1404: 5Ј-GCATTTGCTGATTGCACAGTAATTCTCGAAG- CAATGCTGGAATGCC-3Ј and 5Ј-GGCATTCCAGCATTGCTTCGAGA- ATTACTGTGCAATCAGCAAATGC-3Ј; C1400A/E1401A, 5Ј-GATTGC- ACAGTAATTCTCGCTGCACACAGGATAGAAGCAATGC-3Ј and 5Ј-G- CATTGCTTCTATCCTGTGTGCAGCGAGAATTACTGTGCAATC-3Ј; H1402A/R1403A, 5Ј-CAGTAATTCTCTGTGAAGCCGCGATAGAAGC- AATGCTGGAATGCC-3Ј and 5Ј-GGCATTCCAGCATTGCTTCTATCG- CGGCTTCACAGAGAATTAC TG-3Ј; I1404A/E1405A, 5Ј-CTCTGTGA- ACACAGGGCAGCAGCAATGCTGGAATGCCAAC-3Ј and 5Ј-GTTGGC- ATTCCAGCATTGCTGCTGCCCTGTGTTCACAGAG-3Ј, Q1390A/A- 1391A/F1392A/A1393A/D1394A, 5Ј-GAAGAACTCTAAAAGCAGCAG- CTGCTGCTTGCACAGTAATTCTC-3Ј and 5Ј-GAGAATTACTGTGCA- AGCAGCAGCTGCTGCTTTTAGAGTTCTTC-3Ј.
X
ABCC7 p.Arg1403Ala 11022033:40:1842
status: NEWX
ABCC7 p.Arg1403Ala 11022033:40:2315
status: NEW[hide] Aggregation of misfolded proteins can be a selecti... J Biol Chem. 2002 Sep 13;277(37):34462-70. Epub 2002 Jun 25. Milewski MI, Mickle JE, Forrest JK, Stanton BA, Cutting GR
Aggregation of misfolded proteins can be a selective process dependent upon peptide composition.
J Biol Chem. 2002 Sep 13;277(37):34462-70. Epub 2002 Jun 25., 2002-09-13 [PMID:12084728]
Abstract [show]
Intracellular aggregation of misfolded proteins is observed in a number of human diseases, in particular, neurologic disorders in which expanded tracts of polyglutamine residues play a central role. A variety of other proteins are prone to aggregation when mutated, indicating that this process is a common pathologic mechanism for inherited disorders. However, little is known about the relationship between the sequence of aggregating peptides and the specificity of intracellular accumulation. Here we demonstrate that substitution of two residues eliminates aggregation of a 111-amino acid peptide derived from the C-terminal portion of the cystic fibrosis transmembrane conductance regulator (CFTR). We also show that fusion to a reporter protein considerably alters the subcellular distribution of aggregating peptide. When fused to green fluorescent protein, the peptide containing amino acids 1370-1480 of CFTR accumulates in large perinuclear or nuclear aggregates. The same CFTR fragment devoid of green fluorescent protein localizes predominantly to discrete accumulations associated with mitochondria. Importantly, both types of accumulation are dependent on the presence of the same two amino acids within the CFTR sequence. Co-expression studies show that both CFTR-derived proteins can co-localize in large cytoplasmic/nuclear aggregates. However, neither CFTR construct accumulates in intracellular inclusions formed by N-terminal fragment of huntingtin. In addition to unique accumulation patterns, each aggregating peptide shows differences in association with chaperone proteins. Thus, our results indicate that the process of intracellular aggregation can be a selective process determined by the composition of the aggregating peptides.
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No. Sentence Comment
90 Only 25% (52/208) of cells expressing GFP fusion protein bearing single H1402A substitution and 27% (41/153) of R1403A mutants showed intracellular protein accumulation, whereas the corresponding number for the wild type sequence was 91% (132/145).
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ABCC7 p.Arg1403Ala 12084728:90:112
status: NEW93 Cells expressing the double mutant H1402A,R1403A showed no aggregation at all (0/108).
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ABCC7 p.Arg1403Ala 12084728:93:42
status: NEW97 However, the introduction of the ⌬ag deletion or the H1402A,R1403A double mutation significantly decreased the amount of the fusion protein in the insoluble fraction to approximately 31 and 36%, respectively.
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ABCC7 p.Arg1403Ala 12084728:97:68
status: NEW108 B-E, the effect of different mutations within the ag region, including ⌬ag (B), V1397E (C), T1396A (D), and double mutation H1402A,R1403A (E) on the aggregation of C terminus of CFTR fused to GFP (shown in green) in transiently transfected human airway epithelial (IB3-1) cells.
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ABCC7 p.Arg1403Ala 12084728:108:138
status: NEW120 Additionally, the double mutation H1402A,R1403A prevented aggregation of HA-tagged CFTR C terminus (Fig. 2E).
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ABCC7 p.Arg1403Ala 12084728:120:41
status: NEW146 E, lack of aggregation of HA-CFTR 1370-1480 construct carrying the H1402A and R1403A mutations.
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ABCC7 p.Arg1403Ala 12084728:146:78
status: NEW160 Similarly, the non-aggregating GFP-CFTR 1370-1480 H1402A,R1403A construct was included in giant aggregates formed by HA-CFTR 1370-1480 (Fig. 4D).
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ABCC7 p.Arg1403Ala 12084728:160:57
status: NEW195 C, colocalization of the aggregating GFP-CFTR 1370-1480 construct (green) and the non-aggregating HA-CFTR 1370-1480 H1402A,R1403A construct (red) in giant aggregates.
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ABCC7 p.Arg1403Ala 12084728:195:123
status: NEW196 D, co-localization of the non-aggregating GFP-CFTR 1370-1480 H1402A,R1403A construct (green) with the aggregating HA-CFTR 1370-1480 construct (red) in giant aggregates.
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ABCC7 p.Arg1403Ala 12084728:196:68
status: NEW[hide] The H-loop in the second nucleotide-binding domain... Cell Physiol Biochem. 2010;25(2-3):169-80. Epub 2010 Jan 12. Kloch M, Milewski M, Nurowska E, Dworakowska B, Cutting GR, Dolowy K
The H-loop in the second nucleotide-binding domain of the cystic fibrosis transmembrane conductance regulator is required for efficient chloride channel closing.
Cell Physiol Biochem. 2010;25(2-3):169-80. Epub 2010 Jan 12., [PMID:20110677]
Abstract [show]
The cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that functions as a cAMP-activated chloride channel. The recent model of CFTR gating predicts that the ATP binding to both nucleotide-binding domains (NBD1 and NBD2) of CFTR is required for the opening of the channel, while the ATP hydrolysis at NBD2 induces subsequent channel closing. In most ABC proteins, efficient hydrolysis of ATP requires the presence of the invariant histidine residue within the H-loop located in the C-terminal part of the NBD. However, the contribution of the corresponding region (H-loop) of NBD2 to the CFTR channel gating has not been examined so far. Here we report that the alanine substitution of the conserved dipeptide HR motif (HR-->AA) in the H-loop of NBD2 leads to prolonged open states of CFTR channel, indicating that the H-loop is required for efficient channel closing. On the other hand, the HR-->AA substitution lead to the substantial decrease of CFTR-mediated current density (pA/pF) in transfected HEK 293 cells, as recorded in the whole-cell patch-clamp analysis. These results suggest that the H-loop of NBD2, apart from being required for CFTR channel closing, may be involved in regulating CFTR trafficking to the cell surface.
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No. Sentence Comment
30 Mutations within the H-loop of NBD2, including the deletion of the entire ag region (Δ1395-1403) and the double alanine substitution H1402A, R1403A (HR→AA), were created in the CFTR-containing pBQ4.7 vector (a gift from J. Rommens and L.-C. Tsui) using the site-directed mutagenesis system "Transformer" (Becton Dickinson).
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ABCC7 p.Arg1403Ala 20110677:30:147
status: NEW[hide] PDZ-binding motifs are unable to ensure correct po... FEBS Lett. 2005 Jan 17;579(2):483-7. Milewski MI, Lopez A, Jurkowska M, Larusch J, Cutting GR
PDZ-binding motifs are unable to ensure correct polarized protein distribution in the absence of additional localization signals.
FEBS Lett. 2005 Jan 17;579(2):483-7., [PMID:15642363]
Abstract [show]
The C-terminal PDZ-binding motifs are required for polarized apical/basolateral localization of many membrane proteins. To determine the specificity of the PDZ-binding motifs in establishing cellular distribution, we utilized a 111-amino acid region from the C-terminus of cystic fibrosis transmembrane conductance regulator (CFTR) that is able to direct apical localization of fused reporter proteins. Substitution of the C-terminal PDZ-binding motif of CFTR with corresponding motifs necessary for basolateral localization of other membrane proteins did not lead to the redistribution of the fusion protein to the basolateral membrane. Instead, some fusion proteins remained localized to the apical membrane, whereas others showed no specific distribution. The specificity of the PDZ-based interactions was substantially increased when specific amino acids located upstream of the classical PDZ-binding motifs were included. However, even the presence of a longer C-terminal motif from a basolateral protein could not ensure basolateral distribution of the fusion protein. Our results indicate that the C-terminal PDZ-binding motifs are not the primary signals for polarized protein distribution, although they are required for targeting and/or stabilization of protein at the given location.
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No. Sentence Comment
25 Also, the introduction of the H1402A and R1403A mutations, reducing the aggregation rate of the CFTR C-terminus, was previously described [9].
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ABCC7 p.Arg1403Ala 15642363:25:41
status: NEW64 A diagram illustrating the structure and subcellular localization of the GFP fusion proteins containing either the full-length CFTR protein or its C-terminal fragment (a.a. 1370-1480) with two alanine substitutions (H1402A and R1403A), eliminating the aggregation of the CFTR C-terminus.
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ABCC7 p.Arg1403Ala 15642363:64:227
status: NEW93 (B) Apical localization of the GFP-CFTR 1370-1480 H1402A, R1403A fusion protein containing the C-terminal M-K-D-T-R-L> motif.
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ABCC7 p.Arg1403Ala 15642363:93:58
status: NEW94 (C) Predominant cytoplasmic distribution of the GFP-CFTR 1370-1480 H1402A, R1403A fusion protein containing the extended Q-K-E-TC-L> PDZ-binding motif of LET23.
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ABCC7 p.Arg1403Ala 15642363:94:75
status: NEW