ABCMdb

ABCC7 p.Cys491Ser

None has been submitted yet.

Publications

PMID: 16442101 [PubMed] Frelet A et al: "Insight in eukaryotic ABC transporter function by mutation analysis."

No. Sentence Comment

307 [138] A455E, P574H cAMP-stimulated apical membrane Cl-currents but current magnitudes were reduced compared to wild-type Electrophysiology of epithelial cells [139] C491S, C1344S, C1355S C491S channels opened almost exclusively to a 3-pS subconductance. Login to comment

PMID: 15272010 [PubMed] Chen EY et al: "The DeltaF508 mutation disrupts packing of the transmembrane segments of the cystic fibrosis transmembrane conductance regulator."

No. Sentence Comment

57 The construction of Cys-less CFTR (C76S/C126S/C225S/C276S/C343S/C491S/C524S/C590S/C592S/C657S/C832S/C866S/C1344S/C1355S/C1395S/C1400S/C1410S/C1458S) was performed using the following cDNA fragments. Login to comment
58 Point mutations C76/126S were generated in sequence in the PstI (bp 1) 3 XbaI (bp 573) fragment; point mutations C225S/C276S/C343S were generated in sequence in the XbaI (bp 573) 3 KpnI (bp 1370) fragment; point mutations C491S/C524S/C590S/C592S/C657S were generated in sequence in the KpnI (bp 1370) 3 ApaI (bp 2333) fragment; point mutations C832S/C866S were generated in sequence in the ApaI (bp 2333) 3 EcoRI (bp 3643) fragment; point mutations C1344S/C1355S/ C1395S/C1400S/C1410S/C1458S were generated in sequence in the EcoRI (bp 3643) 3 XhoI (bp 4560) fragment, the five insert fragments were then ligated and inserted into the PstI and XhoI sites of plasmid vector pMT21. Login to comment

PMID: 16189704 [PubMed] McGinniss MJ et al: "Extensive sequencing of the CFTR gene: lessons learned from the first 157 patient samples."

No. Sentence Comment

7 We ascertained ten novel sequence variants that are potentially disease-associated: two deletions (c.1641AG>T, c.2949_2853delTACTC), seven missense mutations (p.S158T, p.G451V, p.K481E, p.C491S, p.H949L, p.T1036N, p.F1099L), and one complex allele ([p.356_A357del; p.358I]). Login to comment
92 The p.C491S missense mutation is expected to be a benign variant since it was ascertained during single exon sequencing in an unaffected sibling of a CF patient. Login to comment
95 The PolyPhen prediction tool also suggests that p.C491S is a benign variant (Table 7). Login to comment

PMID: 17489851 [PubMed] Tzetis M et al: "Contribution of the CFTR gene, the pancreatic secretory trypsin inhibitor gene (SPINK1) and the cationic trypsinogen gene (PRSS1) to the etiology of recurrent pancreatitis."

No. Sentence Comment

93 a Additional mutations found in the controls: p.R1162L (1.66%), p.D565G (0.47%), p.A120T (0.47%) and 0.24% each for p.R297Q, p.L997F, p.E826K, p.I807M, p.S495Y and p.C491S. Login to comment

PMID: 19754156 [PubMed] Alexander C et al: "Cystic fibrosis transmembrane conductance regulator: using differential reactivity toward channel-permeant and channel-impermeant thiol-reactive probes to test a molecular model for the pore."

No. Sentence Comment

42 The Cys-less CFTR construct (C76S, C126S, C225S, C276S, C343S, C491S, C524S, C590L, C592L, C657S, C832S, C866S, C1344S, C1355S, C1395S, C1400S, C1410S, C1458S) was a gift from Drs. Martin Mense and David Gadsby and was used in their pGEMHE vector previously described (13). Login to comment

PMID: 22326559 [PubMed] Poulou M et al: "Cystic fibrosis genetic counseling difficulties due to the identification of novel mutations in the CFTR gene."

No. Sentence Comment

62 (3) Disease causing Atypical CF 15 14 (13) c.2450GNT p.Gly817Val Benign T 0.31 Path. (4) Polymorphism CBAVD 16 11 (10) c.1472GNC p.Cys491Ser Benign T 0.59 Neut. Login to comment
73 Disruption of ESE motifs/creation of ESS motifs Acc increased (0.44/0.50) Acc gained 0.51 p.V1318G c.3953TNG No change New donor site (48.15/74.98) (+55.73%) if used causes reduction of exon size (-11nt) Donor increased (0.31/0.99) p.K1165T c.3494ANC No change Minor changes No change p.S977C c.2930CNG No change Minor changes No change p.I521F c.1561ANT No change Minor changes No change p.V1212F c.3634GNT No change Disruption of ESE motifs Donor gained 0.36 p.F319V c.955TNG No change Minor changes Acc increased (0.42/0.61) p.G817V c.2450GNT No change Minor changes No change p.C491S c.1472GNC No change Disruption of ESE motifs Acceptor sites created p.I336L c.1006ANC No change Disruption of ESE motifs No change p.F305V c.913TNG Increased score for SRp55 Disruption of ESS motifs Donor gained (0.99) p.S511C c.1532CNG Decreased score for SC35 Disruption of ESS motifs No change p.S1311N c.3932GNA Changes on ss scores Disruption of ESS motifs Donor increased (0.50/0.63) p.L926F c.2778GNT No change Creation of ESS motifs Changes on ss scores p.A1225V c.3674CNT No change Disruption of ESE motif/creation of ESS motif Donor increased (0.72/0.99) p.F533L c.1597TNC No change Creation of ESE motifs/disruption of ESS motifs No change p.Q1209H c.3627ANC No change Minor changes No change c.2490+3ANG No change Minor changes No change c.2909-36TNC Decrease of donor ss Disruption of ESS motifs Acc increased (0.33/0.48)/donor increased (0.74/0.94) c.2909-10TNC Change for SRp55 best hit (3.97/5.16) Disruption of ESS motifs No change c.4137-21GNT No change Disruption of ESE and creation of ESS motifs Acc increased (0.86/0.96) c.1116+4ANT No change WT donor site disrupted Marginal changes on donor ss c.2988+30TNC No change Disruption of ESE motif/creation of ESS motif No change c.1680-27GNA No change Disruption of ESS motifs No change c.2620-24CNG No change Creation of ESE motifs Donor gained 0.96 c.2620-18delT No change New donor ss/changes on ESE and ESS motifs Acc increased (0.41/0.55) c.2658-8CNG Decreased score for acc site Minor changes Marginal changes on ss p.L1227L c.3681ANG No change Changes on ESE and ESS motifs No change p.R1158R c.3472CNA No change Creation of ESE motifs/creation of ESS motifs Marginal changes on ss p.D1275D c.3825TNC No change No change Donor increased (0.56/0.87) p.L346L c.1036CNT No change No change No change Abbreviations: ESE: exonic splicing enhancer, HSF: human splicing finder, ESS: exonic splicing silencer, WT: wild type, Mut: mutant, ss: splicing site. Login to comment
75 Disruption of ESE motifs/creation of ESS motifs Acc increased (0.44/0.50) Acc gained 0.51 p.V1318G c.3953TNG No change New donor site (48.15/74.98) (+55.73%) if used causes reduction of exon size (-11nt) Donor increased (0.31/0.99) p.K1165T c.3494ANC No change Minor changes No change p.S977C c.2930CNG No change Minor changes No change p.I521F c.1561ANT No change Minor changes No change p.V1212F c.3634GNT No change Disruption of ESE motifs Donor gained 0.36 p.F319V c.955TNG No change Minor changes Acc increased (0.42/0.61) p.G817V c.2450GNT No change Minor changes No change p.C491S c.1472GNC No change Disruption of ESE motifs Acceptor sites created p.I336L c.1006ANC No change Disruption of ESE motifs No change p.F305V c.913TNG Increased score for SRp55 Disruption of ESS motifs Donor gained (0.99) p.S511C c.1532CNG Decreased score for SC35 Disruption of ESS motifs No change p.S1311N c.3932GNA Changes on ss scores Disruption of ESS motifs Donor increased (0.50/0.63) p.L926F c.2778GNT No change Creation of ESS motifs Changes on ss scores p.A1225V c.3674CNT No change Disruption of ESE motif/creation of ESS motif Donor increased (0.72/0.99) p.F533L c.1597TNC No change Creation of ESE motifs/disruption of ESS motifs No change p.Q1209H c.3627ANC No change Minor changes No change c.2490+3ANG No change Minor changes No change c.2909-36TNC Decrease of donor ss Disruption of ESS motifs Acc increased (0.33/0.48)/donor increased (0.74/0.94) c.2909-10TNC Change for SRp55 best hit (3.97/5.16) Disruption of ESS motifs No change c.4137-21GNT No change Disruption of ESE and creation of ESS motifs Acc increased (0.86/0.96) c.1116+4ANT No change WT donor site disrupted Marginal changes on donor ss c.2988+30TNC No change Disruption of ESE motif/creation of ESS motif No change c.1680-27GNA No change Disruption of ESS motifs No change c.2620-24CNG No change Creation of ESE motifs Donor gained 0.96 c.2620-18delT No change New donor ss/changes on ESE and ESS motifs Acc increased (0.41/0.55) c.2658-8CNG Decreased score for acc site Minor changes Marginal changes on ss p.L1227L c.3681ANG No change Changes on ESE and ESS motifs No change p.R1158R c.3472CNA No change Creation of ESE motifs/creation of ESS motifs Marginal changes on ss p.D1275D c.3825TNC No change No change Donor increased (0.56/0.87) p.L346L c.1036CNT No change No change No change Abbreviations: ESE: exonic splicing enhancer, HSF: human splicing finder, ESS: exonic splicing silencer, WT: wild type, Mut: mutant, ss: splicing site. Login to comment

PMID: 11867445 [PubMed] Harrington MA et al: "Cysteine residues in the nucleotide binding domains regulate the conductance state of CFTR channels."

No. Sentence Comment

72 C491S, C524S, C1344/1355S, and C491/524S mutants were inserted into a pcDNA3.1 expression vector and transiently transfected into HEK 293 cells using calcium phosphate. Login to comment
91 In inside-out patch clamp recordings, C491S mutant channels show openings to two different conductance levels, with the majority of channel openings to a subconductance of ϳ3 pS (Fig. 2, Table 1). Login to comment
93 The subconductance openings of the C491S mutant are much shorter than the wild-type subconductance, as shown by comparing the dwell time histograms in Fig. 1 C with that in Fig. 2 C. Login to comment
95 Despite this difference, the C491S subconductance openings appeared to be very similar to the wild-type subconductance in current amplitudes and in requirement for phosphorylation. Login to comment
96 As shown in Fig. 3, C491S mutant channels, like the wild-type subconductance, require phosphorylation by PKA for high-probability openings. Login to comment
98 While the C491S single mutation had occasional full-size openings along with subconductance openings, mutating both cysteine residues in NBD1 resulted in a channel that almost never opened to the full-size conductance (Table 1; Fig. 4, A and B). Login to comment
104 As shown by the dwell time histograms in Fig. 4 C, the long component of the open time distributions for the C491/524S and C-QUAD-S mutants is much shorter than the wild-type subconductance, and closer to that of the C491S mutant. Login to comment
105 Mutation of C524 has little effect on channel gating While CFTR channels carrying the C491S mutation either alone or in combination with C524S or C1344/1355S open almost exclusively to a 3 pS subconductance, channels carry- ing only the C524S mutation exhibit conductance similar to wild-type channels. Login to comment
106 Like the wild-type channel, nearly every patch of C524S channels gates to the full-size openings, although, like wild-type channels, subconductance openings do appear (Table 1). Login to comment
117 In the presence of a mixture of ATP and ATP␥S, full-size channels are "locked open": opening in long bursts that can last for minutes (Anderson et al., 1991b; Baukrowitz et al., FIGURE 2 C491S-CFTR channels open most frequently to a 3-pS subconductance. Login to comment
118 (A) One-minute sample traces of inside-out patches from HEK 293 cells expressing the C491S mutant of CFTR showing the two modes of gating of the channel. Login to comment
122 (C) Open dwell time histogram of 6000 opening and closing events of C491S channels in nine separate patches. Login to comment
124 (D) Current-voltage relationship of subconductance channels from C491S CFTR. Login to comment
131 Two other mutants (C491S, C491/524S) were tested for the effect of ATP␥S on subconductance openings (Fig. 5 C). Login to comment
132 Both the C491S and C491/524S mutants have open dwell times that are shorter than wild-type subconductance, and these dwell times were not significantly altered by the presence of ATP␥S (p Ͼ 0.05; Kolmogorov-Smirnov). Login to comment
134 This is in contrast FIGURE 3 Increased phosphorylation by PKA increases the frequency of subconductance openings in C491S patches. Login to comment
135 (A) One-minute sample traces of an inside-out patch from an HEK 293 cell expressing C491S mutant CFTR with and without the addition of PKA. Login to comment
138 The patch was held at 75 mV. (B) Graph of open probability versus time for the patch shown in A. TABLE 1 Mutations of cysteine residues in NBD1 increase the proportion of patches with subconductance openings, while mutations of cysteines in NBD2 decrease the proportion of patches with subconductance openings Mutation Patches with Subconductance Patches with Full Size Total Patches Percent with Subconductance Percent with Full Size Wild-type 34 49 49 69 100 C491S 16 5 18 89 28 C491/524S 10 1 13 77 8 C524S 2 5 6 33 83 C1344/1355S 8 30 30 27 100 C-QUAD-S 19 0 30 63 0 The number of inside-out patches containing frequent full-size and/or subconductance channel openings were counted. Login to comment
143 Moreover, the presence of ATP␥S shortens the open dwell time of wild-type channels to approximately that of channels containing the C491S mutant, suggesting that the altered behavior of the C491S channel may be related to changes in ATP hydrolysis at NBF1. Login to comment
181 In some cases the channel transitions to the subconductance without appearing to close, while sometimes it opens directly to the subconductance state and then transitions to the full-size channel. This type of rapid transition between full-size and subconductance was not observed in recordings of wild-type channels in reducing conditions, nor in the C491S, C491/524S, or C- QUAD-S mutants in which the subconductance state makes up the majority of channel openings observed. Login to comment
183 Effect of mutation of C1344 and C1355 on subconductance frequency and redox sensitivity Mutation of cysteine residues in the second nucleotide binding domain by themselves had much less of an effect on channel gating than the C491S mutation. Login to comment
206 The 3-pS subconductance of channels containing the C491S mutant alone or with other cysteine mutations was similar to the subconductance observed in recordings from patches containing wild-type CFTR channels, although with a shortened open dwell time. Login to comment
208 The effect of the C491S mutation on control of channel conductance (enriching for subconductance at the expense of full-size openings) was unexpected; however, the specificity of this mutation for that effect is supported by several pieces of data. Login to comment
221 With channels containing the C491S mutation opening almost exclusively to a subconductance with gating properties very different from the full-size channel, it is difficult to directly relate the effect of redox potential on gating kinetics of the mutants compared to the wild-type channel. Login to comment
244 Our data suggest that subconductance opening may be related to a decreased hydrolysis by the first nucleotide binding domain because mutation of C491S in NBD1 results in an increase in subconductance frequency at the expense of the full-size openings. Recently published work with truncation mutants of CFTR has suggested that the CFTR channel might be "double-barreled," with one pore producing the full-size conductance and a second, independently gated pore producing a 3-4-pS subconductance (Yue et al., 2000). Login to comment
256 The frequency of subconductance openings in CFTR channels with the C491S mutation may mean that this region of the molecule is important for the type of intermolecular interactions observed by other groups to be important for high-frequency opening of the full-size channel. Login to comment
73 C491S, C524S, C1344/1355S, and C491/524S mutants were inserted into a pcDNA3.1 expression vector and transiently transfected into HEK 293 cells using calcium phosphate. Login to comment
92 In inside-out patch clamp recordings, C491S mutant channels show openings to two different conductance levels, with the majority of channel openings to a subconductance of b03;3 pS (Fig. 2, Table 1). Login to comment
94 The subconductance openings of the C491S mutant are much shorter than the wild-type subconductance, as shown by comparing the dwell time histograms in Fig. 1 C with that in Fig. 2 C. Login to comment
97 As shown in Fig. 3, C491S mutant channels, like the wild-type subconductance, require phosphorylation by PKA for high-probability openings. Login to comment
99 While the C491S single mutation had occasional full-size openings along with subconductance openings, mutating both cysteine residues in NBD1 resulted in a channel that almost never opened to the full-size conductance (Table 1; Fig. 4, A and B). Login to comment
119 (A) One-minute sample traces of inside-out patches from HEK 293 cells expressing the C491S mutant of CFTR showing the two modes of gating of the channel. Login to comment
123 (C) Open dwell time histogram of 6000 opening and closing events of C491S channels in nine separate patches. Login to comment
125 (D) Current-voltage relationship of subconductance channels from C491S CFTR. Login to comment
133 Both the C491S and C491/524S mutants have open dwell times that are shorter than wild-type subconductance, and these dwell times were not significantly altered by the presence of ATPॹS (p b0e; 0.05; Kolmogorov-Smirnov). Login to comment
136 (A) One-minute sample traces of an inside-out patch from an HEK 293 cell expressing C491S mutant CFTR with and without the addition of PKA. Login to comment
140 TABLE 1 Mutations of cysteine residues in NBD1 increase the proportion of patches with subconductance openings, while mutations of cysteines in NBD2 decrease the proportion of patches with subconductance openings Mutation Patches with Subconductance Patches with Full Size Total Patches Percent with Subconductance Percent with Full Size Wild-type 34 49 49 69 100 C491S 16 5 18 89 28 C491/524S 10 1 13 77 8 C524S 2 5 6 33 83 C1344/1355S 8 30 30 27 100 C-QUAD-S 19 0 30 63 0 The number of inside-out patches containing frequent full-size and/or subconductance channel openings were counted. Login to comment
145 Moreover, the presence of ATPॹS shortens the open dwell time of wild-type channels to approximately that of channels containing the C491S mutant, suggesting that the altered behavior of the C491S channel may be related to changes in ATP hydrolysis at NBF1. Login to comment
185 Effect of mutation of C1344 and C1355 on subconductance frequency and redox sensitivity Mutation of cysteine residues in the second nucleotide binding domain by themselves had much less of an effect on channel gating than the C491S mutation. Login to comment
210 The effect of the C491S mutation on control of channel conductance (enriching for subconductance at the expense of full-size openings) was unexpected; however, the specificity of this mutation for that effect is supported by several pieces of data. Login to comment
223 With channels containing the C491S mutation opening almost exclusively to a subconductance with gating properties very different from the full-size channel, it is difficult to directly relate the effect of redox potential on gating kinetics of the mutants compared to the wild-type channel. Login to comment
247 Our data suggest that subconductance opening may be related to a decreased hydrolysis by the first nucleotide binding domain because mutation of C491S in NBD1 results in an increase in subconductance frequency at the expense of the full-size openings. Recently published work with truncation mutants of CFTR has suggested that the CFTR channel might be "double-barreled," with one pore producing the full-size conductance and a second, independently gated pore producing a 3-4-pS subconductance (Yue et al., 2000). Login to comment
259 The frequency of subconductance openings in CFTR channels with the C491S mutation may mean that this region of the molecule is important for the type of intermolecular interactions observed by other groups to be important for high-frequency opening of the full-size channel. It is conceivable that the cysteine residues in NBD1, particularly C491, could be important in the intermolecular associations that stabilize a dimer of the channel and allow it to produce full-size openings. Login to comment

PMID: 16766608 [PubMed] Serrano JR et al: "CFTR: Ligand exchange between a permeant anion ([Au(CN)2]-) and an engineered cysteine (T338C) blocks the pore."

No. Sentence Comment

23 MATERIALS AND METHODS Mutagenesis and in vitro transcription The Cys-less CFTR construct (C76S, C126S, C225S, C276S, C343S, C491S, C524S, C590L, C592L, C657S, C832S, C866S, C1344S, C1355S, C1395S, C1400S, C1410S, C1458S) was a gift from Drs. Martin Mense and Submitted December 28, 2005, and accepted for publication May 19, 2006. Login to comment

PMID: 26149808 [PubMed] Chong PA et al: "Deletion of Phenylalanine 508 in the First Nucleotide-binding Domain of the Cystic Fibrosis Transmembrane Conductance Regulator Increases Conformational Exchange and Inhibits Dimerization."

No. Sentence Comment

288 Of note, we were not able to express or purify NBD1 using the previously published mutations C491S, C524S, C590V, and C592V (12), probably because these mutations destabilize NBD1. Login to comment