ABCMdb

ABCC7 p.Ser1248Phe

None has been submitted yet.

Publications

PMID: 15623556 [PubMed] Berger AL et al: "Normal gating of CFTR requires ATP binding to both nucleotide-binding domains and hydrolysis at the second nucleotide-binding domain."

No. Sentence Comment

103 The A462F mutation prevented labeling of NBD1; S1248F blocked NBD2 labeling; and the double mutant A462F͞S1248F abolished labeling at both NBDs. Login to comment
110 The S1248F mutation reduced Po by prolonging the interburst interval without altering burst duration (Fig. 2 A and C). Login to comment
113 In addition, correspondence of the S1248F and NEM-modified S1248C data indicate that the gating effects of the S1248F substitution were not due to misfolding that might have occurred during channel biosynthesis. Login to comment
127 (A) Examples of single-channel recordings for WT and S1248F CFTR. Login to comment
140 The finding that channels unable to bind nucleotide at NBD2 (S1248F and NEM-modified S1248C) had a normal burst duration suggested that the prolonged burst duration of K1250A (16-18, 20, 21) arose when ATP bound NBD2 but then did not undergo hydrolysis. Login to comment
143 However, when we modified NBD2 with NEM to prevent nucleotide binding, burst duration fell into the range of WT or S1248F channels (compare Figs. Login to comment
172 The S1248F mutation will block NBD2 ATP binding, thereby limiting ATP interactions to NBD1, and the K464A mutation will prevent NBD1 ATP hydrolysis. Login to comment
174 However with ATP, the Po, interburst interval, and burst duration were similar to those obtained with the S1248F mutation alone (Figs. Login to comment
203 Second, the K464A͞S1248F mutant behaved similarly to the S1248F single mutation. Login to comment
221 Our results argue against this explanation because blocking NBD2 ATP binding (with either the S1248F mutation or NEM-modification of S1248C) did not prolong burst duration. Login to comment
233 However, it seems surprising that blocking ATP binding to NBD2 (with the S1248F mutation or the NEM-modified S1248C mutation) did not change the rate of channel closure compared to WT. Login to comment
250 However, that is not what we observed; the A462F mutation completely blocked NBD1 labeling, and S1248F blocked NBD2 labeling. Login to comment
253 Nevertheless, we did find that the A462F mutation in NBD1 diminished labeling of NBD2 and that S1248F slightly reduced NBD1 labeling. Login to comment

PMID: 22948143 [PubMed] Randak CO et al: "Demonstration of Phosphoryl Group Transfer Indicates That the ATP-binding Cassette (ABC) Transporter Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Exhibits Adenylate Kinase Activity."

No. Sentence Comment

56 CFTR Adenylate Kinase Assay-Membranes containing either 30 ␮g of protein (from cells expressing wild-type CFTR) or 90 ␮g of protein (from cells expressing S1248F CFTR) were incubated gently shaking with nonradioactive 8- or 2-N3-AMP (at concentrations given in the figure legends), radioactive [␥-32 P]GTP (30 ␮Ci, 6000 Ci/mmol), 20 mM Hepes (pH 7.5), 50 mM NaCl, 3 mM MgCl2, and 1 mM Tricine (pH 7.6) for 5 min at 37 °C in a total volume of 30 ␮l followed by UV irradiation for 30 s (302 nm, 8-watt lamp) at a distance of 5 cm. Login to comment
137 We chose a phenylalanine substitution for serine at position 1248 (S1248F) in the phosphate-binding loop of ATP-binding site 2. Login to comment
151 In lane 6, membranes containing 90 ␮g of protein from S1248F CFTR-expressing HeLa cells were employed. Login to comment
161 30 ␮g (control membranes and membranes with wild-type CFTR, lanes 1-3) and 90 ␮g (membranes with S1248F CFTR, lane 4) of protein were used. Login to comment
163 When we incubated membranes containing S1248F CFTR with [␥-32 P]GTP and nonradioactive N3-AMP, followed by UV irradiation, we found very little labeling (Fig. 4A, lane 6). Login to comment
178 Substituting a phenylalanine into the phosphate-binding loop of NBD2 (the S1248F mutation) interfered with labeling. Login to comment
180 Previous observations support the interpretation that the S1248F mutation disrupted adenylate kinase activity. Login to comment
181 1) A study characterizing the gating characteristics and the interaction of ATP with S1248F CFTR found that this mutation interfered with the interaction of nucleotides at ATP-binding site 2. Login to comment
57 CFTR Adenylate Kinase Assay-Membranes containing either 30 òe;g of protein (from cells expressing wild-type CFTR) or 90 òe;g of protein (from cells expressing S1248F CFTR) were incubated gently shaking with nonradioactive 8- or 2-N3-AMP (at concentrations given in the figure legends), radioactive [ॹ-32 P]GTP (30 òe;Ci, 6000 Ci/mmol), 20 mM Hepes (pH 7.5), 50 mM NaCl, 3 mM MgCl2, and 1 mM Tricine (pH 7.6) for 5 min at 37 &#b0;C in a total volume of 30 òe;l followed by UV irradiation for 30 s (302 nm, 8-watt lamp) at a distance of 5 cm. Login to comment
136 We chose a phenylalanine substitution for serine at position 1248 (S1248F) in the phosphate-binding loop of ATP-binding site 2. Login to comment
150 In lane 6, membranes containing 90 òe;g of protein from S1248F CFTR-expressing HeLa cells were employed. Login to comment
160 30 òe;g (control membranes and membranes with wild-type CFTR, lanes 1-3) and 90 òe;g (membranes with S1248F CFTR, lane 4) of protein were used. Login to comment
162 When we incubated membranes containing S1248F CFTR with [ॹ-32 P]GTP and nonradioactive N3-AMP, followed by UV irradiation, we found very little labeling (Fig. 4A, lane 6). Login to comment

PMID: 23921386 [PubMed] Randak CO et al: "ATP and AMP mutually influence their interaction with the ATP-binding cassette (ABC) adenylate kinase cystic fibrosis transmembrane conductance regulator (CFTR) at separate binding sites."

No. Sentence Comment

239 Substituting serine at position 1248 in NBD2 with phenylalanine (S1248F mutation; Fig. 9C, left) abolished nucleotide interaction with ATP-binding site 2. Login to comment
241 The A462F, but not the S1248F mutation interfered with processing and trafficking to the cell membrane (supplemental Fig. S1), and hence, the number of channels in excised membrane patches was small; therefore, we quantified channel activity as NPo. Login to comment
243 On the other hand, Ap5A had no effect on ATP-dependent current of S1248F CFTR (Fig. 9C, middle and right). Login to comment
270 C, left, model of S1248F CFTR. Login to comment
272 Middle, current recording (100 ms averages) from an excised inside-out membrane patch containing multiple S1248F CFTR channels. ATP and Ap5A were present during the times and at the concentrations indicated by bars. Login to comment
273 ATP was added together with PKA catalytic subunit. Holding voltage was afa;80 mV. Right, S1248F CFTR Clafa; current before and after adding 1 mM Ap5A. Experiments were performed as shown in the middle panel with 0.3 mM ATP and PKA present. Login to comment
301 (a) The S1248F mutation, which interfered with the binding of ATP to site 2 (15) and abolished the inhibition of CFTR current in the presence of Ap5A in our study (Fig. 9C), also disrupted CFTR adenylate kinase activity (22). Login to comment

PMID: 25887396 [PubMed] Dong Q et al: "Mutating the Conserved Q-loop Glutamine 1291 Selectively Disrupts Adenylate Kinase-dependent Channel Gating of the ATP-binding Cassette (ABC) Adenylate Kinase Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) and Reduces Channel Function in Primary Human Airway Epithelia."

No. Sentence Comment

165 As a positive control, we mutated serine 1248 to phenylalanine (S1248F) in Q1291F CFTR.TheS1248FmutationpreventstheinteractionofATPwith ATP-bindingsite2(48).Asanticipated,theopenprobabilityofthe double mutant was markedly reduced, mainly due to interburst closed times that were significantly longer than those of wild-type andQ1291FCFTR(comparebar6withbars1and5inthetopand bottom panels of Fig. 5B). Login to comment
207 Holding voltage was afa;80 mV for wild-type, Q1291W, and Q1291Y CFTR and afa;60 mV for Q1291H, Q1291F, and Q1291F/S1248F CFTR. Login to comment