ABCMdb

ABCC7 p.Ser573Glu

None has been submitted yet.

Publications

PMID: 12508051 [PubMed] Vergani P et al: "On the mechanism of MgATP-dependent gating of CFTR Cl- channels."

No. Sentence Comment

154 The mean closing rate from bursts was not substantially altered by these NBD1 mutations (compare Fig. 5 E and Table I): for D572N, rOC(5 mM MgATP ϩ PKA) ϭ 1.4 Ϯ 0.2 s-1 (n ϭ 9), and rOC(5 mM MgATP) ϭ 3.1 Ϯ 0.6 s-1 (n ϭ 3); for S573E, rOC(5 mM MgATP ϩ PKA) ϭ 2.2 Ϯ 0.3 s-1 (n ϭ 7). Login to comment
166 For S573E channels, rCO(5 mM MgATP ϩ PKA) ϭ 0.9 Ϯ 0.2 s-1 (n ϭ 7, total estimate) and 1 Ϯ 0.2 s-1 (n ϭ 4, best estimate). Login to comment
183 Patches contained one WT (A), K464A (B), or S573E (D) channel, or more than one D572N (C) channel. Login to comment
184 (E) Summary of mean (ϮSEM) ␶b values at 5 mM MgATP and 300 nM PKA (n ϭ 30, 21, 9, and 7 for WT, K464A, D572N, and S573E, respectively). Login to comment

PMID: 16989640 [PubMed] Stratford FL et al: "The Walker B motif of the second nucleotide-binding domain (NBD2) of CFTR plays a key role in ATPase activity by the NBD1-NBD2 heterodimer."

No. Sentence Comment

40 For S573E the following primers were used: reverse primer 5 -CTAGGTATCCAAAAGGCTCGTCTAATAAATACAA- AT-3 and forward primer 5 -ATTTGTATTTATTAGACGAGC- CTTTTGGATACCTAG-3 . Login to comment
77 The dose-response of TNP-ATP binding to the mutant NBDs: S573E and E1371Q was analysed using a single site binding algorithm and curve fitting by GraphPad Prism to determine Kd values. Login to comment
84 Similarly, the Walker B mutant proteins (S573E) and (E1371Q) bearing polyhistidine tags and an HA tag, in the case of E1371Q, were expressed in Sf9 cells using the baculovirus system (Figure 1B). Login to comment
86 Both the purified NBD1 mutant (S573E) and purified NBD2 mutant (E1371Q) bind the ATP analogue, TNP-ATP, with low micromolar affinities (Figure 2), similar to the affinities previously reported for wild-type NBD1 (11.6 µM) and NBD2 (10.6 µM) [14]. Login to comment

PMID: 22265409 [PubMed] Mendoza JL et al: "Requirements for efficient correction of DeltaF508 CFTR revealed by analyses of evolved sequences."

No. Sentence Comment

70 Two 508-coupled mutants, D529F and S573E, dramatically increased the relative folding yield of NBD1 by 3.07- &#b1; 0.13-fold and 1.85- &#b1; 0.06-fold, respectively, a level comparable to that of the second-site suppressors identified in the STE6 chimera screen (Figure 3A). Login to comment
71 Thermal denaturation of purified D529F and S573E NBD1 revealed that S573E raised the Tm by 2 C-5 C relative to wild-type depending on ATP concentration (Figure S4B), indicating the improved folding yield in the complementation assay could be accounted for by increased stability of the native state (Figure 3B). Login to comment
108 The same two mutations that improved NBD1 folding yield, D529F and S573E, increased the maturation efficiency of CFTR (Figure 4A, yellow and magenta bars, respectively). Login to comment
115 Top 20 508-Coupled Positions within NBD1 Using Four Independent Statistical Methods ELSC McBASC OMES SCA 435 453 460 S466T S466T 468 470 472 473 473 474 474 L475Y L475Y L475Y F490L F490L W496V W496V W496V W496V 503 505 507 509 512 513 Y517I Y517I Y517I Y517I 520 520 521 C524A C524A C524A C524A L526A L526A L526A L526A D529F D529F D529F D529F D537F D537F 543 Y563V Y563V Y563V Y563V A566P A566P 569 569 S573E P574A P574A P574A F575T F575T 578 582 E583G E583G 587 591 595 598 602 604 604 604 H609T 617 630 630 640 The alignment of 493 sequences was used to calculate pairwise coupling scores using each method (ELSC, SCA, McBASC, and OMES). Login to comment
120 Interestingly, the two suppressors identified by the coupling analysis, D529F and S573E, had much more modest effects on NBD1 yield in the presence of the DF508 mutant as might be expected. Login to comment
127 The surface view A B I539T G550E R553M R555K 3M WT F S466T L475Y F490L W496V Y517I C524A L526A D529F D537F Y563V A566P S573E P574A F575T E583G H609T 0 1 2 3 Relative Yield NBD1 ( -gal.) 25 30 35 40 45 0.0 0.5 1.0 Temperature (C ) Relative Turbitity 0 1 2 3 4 -5 0 5 10 WT F I539T I539T F S573E R555K D529F Relative Yield NBD1 ( -gal.) Tm Figure 3. Login to comment
129 All mutations altered the relative yield relative to wild-type NBD1 as shown in the bar chart (&#b1; standard error of the mean [SEM], n = 9 except for WT, DF508, D529F, and S573E where n = 18). Login to comment
130 Two of the 508-coupled positions, D529F and S573E, increase the yield of NBD1. Login to comment
132 (B) Thermal denaturation of 5 mM purified recombinant WT, D529F, and S573E NBD1 in the presence of 2 mM ATP (left panel). Login to comment
134 S573E (magenta circles) increased the melting temperature 2 C. Login to comment
166 B C A B 0 1 2 3 0 1 2 Relative Yield NBD1 (b2;-gal.) Relative Yield CFTR (ELISA) WT ࢞F WT ƊF S466T L475Y F490L W496V Y517I C524A L526A D529F D537F Y563V A566P S573E P574A F575T E583G H609T 0 1 2 Relative Yield CFTR (ELISA) I539T G550E R553M R555K 3M Figure 4. Login to comment
168 (A) The efficiency of full-length CFTR maturation was determined by ELISA (&#b1;SEM, n = 6 for WT, DF508, D529F, and S573E, n = 3 for other mutants). Login to comment
171 The two 508-coupled positions that increased NBD1 folding yield, D529F and S573E, also increased the maturation yield of CFTR (yellow and magenta bars, respectively). Login to comment
173 The two F508- coupled position mutations, D529F and S573E, are colored in yellow and magenta circles, respectively. Login to comment
185 Previously identified second-site suppressor (I539T, G550E, R553M, R555K, and 3M) but not the 508-coupled mutants (D529F and S573E) increase the yield of DF508 NBD1. Login to comment
190 Identified in this study, D529F and S573E improve folding of NBD1 in isolation and maturation of full-length CFTR in the wild-type background. Login to comment
233 D529F, S573E, and R555K mutations of human wild-type NBD1 were expressed and purified as previously described (Thibodeau et al., 2005). Login to comment