ABCC7 p.Pro759Ala

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PMID: 12054472 [PubMed] Tan AL et al: "Biochemical implications of sequence comparisons of the cystic fibrosis transmembrane conductance regulator."
No. Sentence Comment
167 The mutation P759A results in a longer mean pore open lifetime and a higher chloride efflux [40].
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ABCC7 p.Pro759Ala 12054472:167:13
status: NEW
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169 We also note that Pro759 is adjacent to the region 760-783 and perhaps contributes to the proposed inhibitory conformation in mammals and amphibians; this is corroborated by observation on the effect of P759A.
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ABCC7 p.Pro759Ala 12054472:169:203
status: NEW
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PMID: 10692317 [PubMed] Xie J et al: "Conformation, independent of charge, in the R domain affects cystic fibrosis transmembrane conductance regulator channel openings."
No. Sentence Comment
33 Site-specific mutations were constructed following the manufacturer`s instructions using the following three mutagenesis oligonucleotides (showed 5Ј to 3Ј, with mutated base underlined): P740A, G CTC AGA ATC TGC TAC TAA GGA CAG C (RsaI enzyme restriction site is destroyed); P750A, G GCT GAT GCG AGC CAG TAT CGC CTC (BsrI site is created); P759A/T757S, C CTG AAG CGT GGC CGG AGA GCT GAT (BsaHI site is created and BsrI site is destroyed).
X
ABCC7 p.Pro759Ala 10692317:33:352
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38 Data presented in this paper for mutants containing P759A mutation are for constructs that also contain the unexpected T757S mutation.
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ABCC7 p.Pro759Ala 10692317:38:52
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57 Vesicle preparation Six 75 cm2 flasks of 293 HEK cells transfected with either pCEP4(WT) or CFTR mutants (P3A, P2A, P740A, P750A, P759A) vectors were harvested and lysed following the procedure described previously (Xie et al., 1995, 1996).
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ABCC7 p.Pro759Ala 10692317:57:130
status: NEW
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64 In experiments with WT, P3A, P2A, P740A, P750, and P759A, cis solution also contains 50 units/ml cAMP-dependent protein kinase A catalytic subunit.
X
ABCC7 p.Pro759Ala 10692317:64:51
status: NEW
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147 All three proline mutations are involved in the enhanced activity of the P3A CFTR channel To investigate which proline is critical for the increased open probability of the P3A channel, we constructed all three single proline mutants (P740A, P750A, P759A) and incorporated each of them into the planar lipid bilayer.
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ABCC7 p.Pro759Ala 10692317:147:249
status: NEW
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149 Mutants P740A and P759A CFTR have similar open probability to WT CFTR, whereas P750A CFTR has significantly increased Po.
X
ABCC7 p.Pro759Ala 10692317:149:18
status: NEW
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164 P759A CFTR shows no increase in open probability compared to the wild type, but its mean open life time is significantly longer than that of the wild-type CFTR.
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ABCC7 p.Pro759Ala 10692317:164:0
status: NEW
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165 To test whether mutation of prolines 750 and 759, but not proline 740, would have even higher Po than P3A by combining the increased channel opening of P750A with the increased open life time of P759A, the double proline mutant P2A CFTR (P750A/P759A) was made.
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ABCC7 p.Pro759Ala 10692317:165:0
status: NEW
X
ABCC7 p.Pro759Ala 10692317:165:195
status: NEW
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ABCC7 p.Pro759Ala 10692317:165:244
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167 P2A has an increased mean open life time, like the P759A mutant, but its Po is not significant higher than wild-type CFTR (p ϭ 0.46).
X
ABCC7 p.Pro759Ala 10692317:167:51
status: NEW
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196 The double (P2A; Fig. 10, lane 4) or single proline mutants (P740A, P750A, P759A; Fig. 10, lanes 5-7, respectively) were also fully glycosylated to a similar extent.
X
ABCC7 p.Pro759Ala 10692317:196:75
status: NEW
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204 (B) Representative single channel currents through a double proline mutant (P2A: P750A/P759A) are plotted.
X
ABCC7 p.Pro759Ala 10692317:204:87
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229 The averaged values were Po ϭ 0.204 Ϯ 0.036 (WT), 0.438 Ϯ 0.029 (WT ϩ CyP (Cyclophilin A)), 0.577 Ϯ 0.090 (P3A CFTR), 0.161 Ϯ 0.018 (P740A), 0.426 Ϯ 0.030 (P750A), 0.166 Ϯ 0.034 (P759A), 0.272 Ϯ 0.059 (P2A), respectively.
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ABCC7 p.Pro759Ala 10692317:229:227
status: NEW
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237 It is also FIGURE 10 Heterologous expression of wild-type CFTR and proline mutants in 293 HEK cells: 293 HEK cells transfected with pCEP4(WT), pCEP4(P3A), pCEP4(P2A), pCEP4(P740A), pCEP4(P750A), or pCEP4(P759A) were immunoprecipitated and blotted as described in the Materials and Methods; mAb24-1 that recognizes the C-terminus of CFTR was used in the immunoprecipitation/Western blot.
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ABCC7 p.Pro759Ala 10692317:237:204
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239 Lane 1 (UNT): untransfected 293 HEK cells; lane 2: WT CFTR expressing cells; lanes 3-7: P3A, P2A, P740A, P750A, and P759A CFTR expressing cells, respectively.
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ABCC7 p.Pro759Ala 10692317:239:116
status: NEW
X
ABCC7 p.Pro759Ala 10692317:239:204
status: NEW
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34 Site-specific mutations were constructed following the manufacturer`s instructions using the following three mutagenesis oligonucleotides (showed 5b18; to 3b18;, with mutated base underlined): P740A, G CTC AGA ATC TGC TAC TAA GGA CAG C (RsaI enzyme restriction site is destroyed); P750A, G GCT GAT GCG AGC CAG TAT CGC CTC (BsrI site is created); P759A/T757S, C CTG AAG CGT GGC CGG AGA GCT GAT (BsaHI site is created and BsrI site is destroyed).
X
ABCC7 p.Pro759Ala 10692317:34:352
status: NEW
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39 Data presented in this paper for mutants containing P759A mutation are for constructs that also contain the unexpected T757S mutation.
X
ABCC7 p.Pro759Ala 10692317:39:52
status: NEW
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58 Vesicle preparation Six 75 cm2 flasks of 293 HEK cells transfected with either pCEP4(WT) or CFTR mutants (P3A, P2A, P740A, P750A, P759A) vectors were harvested and lysed following the procedure described previously (Xie et al., 1995, 1996).
X
ABCC7 p.Pro759Ala 10692317:58:130
status: NEW
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65 In experiments with WT, P3A, P2A, P740A, P750, and P759A, cis solution also contains 50 units/ml cAMP-dependent protein kinase A catalytic subunit.
X
ABCC7 p.Pro759Ala 10692317:65:51
status: NEW
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148 All three proline mutations are involved in the enhanced activity of the P3A CFTR channel To investigate which proline is critical for the increased open probability of the P3A channel, we constructed all three single proline mutants (P740A, P750A, P759A) and incorporated each of them into the planar lipid bilayer.
X
ABCC7 p.Pro759Ala 10692317:148:249
status: NEW
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150 Mutants P740A and P759A CFTR have similar open probability to WT CFTR, whereas P750A CFTR has significantly increased Po.
X
ABCC7 p.Pro759Ala 10692317:150:18
status: NEW
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166 To test whether mutation of prolines 750 and 759, but not proline 740, would have even higher Po than P3A by combining the increased channel opening of P750A with the increased open life time of P759A, the double proline mutant P2A CFTR (P750A/P759A) was made.
X
ABCC7 p.Pro759Ala 10692317:166:195
status: NEW
X
ABCC7 p.Pro759Ala 10692317:166:244
status: NEW
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168 P2A has an increased mean open life time, like the P759A mutant, but its Po is not significant higher than wild-type CFTR (p afd; 0.46).
X
ABCC7 p.Pro759Ala 10692317:168:51
status: NEW
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198 The double (P2A; Fig. 10, lane 4) or single proline mutants (P740A, P750A, P759A; Fig. 10, lanes 5-7, respectively) were also fully glycosylated to a similar extent.
X
ABCC7 p.Pro759Ala 10692317:198:75
status: NEW
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206 (B) Representative single channel currents through a double proline mutant (P2A: P750A/P759A) are plotted.
X
ABCC7 p.Pro759Ala 10692317:206:87
status: NEW
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231 The averaged values were Po afd; 0.204 afe; 0.036 (WT), 0.438 afe; 0.029 (WT af9; CyP (Cyclophilin A)), 0.577 afe; 0.090 (P3A CFTR), 0.161 afe; 0.018 (P740A), 0.426 afe; 0.030 (P750A), 0.166 afe; 0.034 (P759A), 0.272 afe; 0.059 (P2A), respectively.
X
ABCC7 p.Pro759Ala 10692317:231:227
status: NEW
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241 Lane 1 (UNT): untransfected 293 HEK cells; lane 2: WT CFTR expressing cells; lanes 3-7: P3A, P2A, P740A, P750A, and P759A CFTR expressing cells, respectively.
X
ABCC7 p.Pro759Ala 10692317:241:116
status: NEW
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