ABCMdb

ABCC7 p.Ser222Glu

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Publications

PMID: 17516627 [PubMed] Wehbi H et al: "Role of the extracellular loop in the folding of a CFTR transmembrane helical hairpin."

No. Sentence Comment

97 When the changes in TM3/4 WT hairpin migration were compared to changes in overall hairpin helicity, a strong correlation (R ) 0.79) was observed (Figure 5), leading us to propose that increases in non-native R-helix structure within ECL2 might Table 1: Migration Behavior on SDS-PAGE Gels of Single and Double Mutants in the Loop Region of CFTR TM3/4 Constructs % change in apparent MW on SDS-PAGE mutant vs TM3/4 WT in WT loop mutantsa vs TM3/4 V232D in V232D loop mutantsa Pb E217G 6.8 ( 0.7 E217S 11.1 ( 3.4 5.4 ( 1.4 0.056 Q220R 15.2 ( 1.1 Q220G 0.3 ( 0.4 Q220N 2.1 ( 1.3 0.5 ( 0.3 0.108 Q220K 14.1 ( 1.0 Q220W 13.1 ( 1.3 11.5 ( 0.9 0.157 Q220E -11.1 ( 1.1 -4.0 ( 0.3 <0.001 S222G 12.0 ( 2.1 1.1 ( 0.6 0.001 S222E -0.3 ( 2.4 1.3 ( 0.5 0.512 E217G/S222G 12.4 ( 1.9 E217S/S222E 26.1 ( 4.5 averagec 10.4 ( 7.3 4.0 ( 4.2 0.067 a Values are the percentage difference vs TM3/4 WT or TM3/4 V232D migration of SDS-PAGE gels. Login to comment
147 For example, S222E and WT migrate at approximately the same rate, but Q220E moves at -11% vs WT; both S222G and E217G/S222G are at +12%; Q220K, Q220R, and Q220W are each at +13-15%. Login to comment
148 Moreover, in construct E217S/S222E, the net charge is not changed vs WT, yet the mutant runs at +26%. Login to comment

PMID: 19181854 [PubMed] Rath A et al: "Detergent binding explains anomalous SDS-PAGE migration of membrane proteins."

No. Sentence Comment

61 PA/VD and ES/SE denote the P205A/V232D and E217S/S222E hairpins, respectively. Login to comment
62 V232D, V232A, P205S, and Q220W) migrated as WT within statistical significance; 2 were faster (V232D and V232K); and 4 were slower (G228L, E217V, E217F, and E217S/S222E). Login to comment
70 The mutant hairpins ranged in loading levels from 3.4-10 g SDS/g, with V232D binding significantly fewer SDS molecules than WT, and the E217F and E217S/S222E mutants binding significantly more. Login to comment
71 All hairpins load more detergent than do intact cytochrome b5, KcsA, and Glut1 [range 0.7-1.7 g SDS/g protein, (8-10)], and the E217F and E217S/S222E mutants are apparently the highest reported binders among the admittedly limited numbers of membrane proteins evaluated to date. Login to comment
85 Gel shifts, SDS binding, helicity, and column MW of TM3/4 hairpins Hairpin* Gel shift (dMW, %) Bound SDS (g/g) Helicity (MRE X 103)† Column MW (mut-wt, %)‡ V232Dcf -11 Ϯ 2.6 3.4 Ϯ 0.9 -17 Ϯ 1.2 ϩ19 Ϯ 1.5 V232K -10 Ϯ 3.0 3.8 Ϯ 0.6 -16 Ϯ 1.1 ϩ5.6 Ϯ 1.6 A204L -2.2 Ϯ 2.3 6.0 Ϯ 0.7 -18 Ϯ 1.3 ϩ3.4 Ϯ 1.6 P205A/V232Dcf ϩ0.12 Ϯ 5.2 4.7 Ϯ 0.4 -19 Ϯ 2.6 ϩ21 Ϯ 0.6 WT ϩ0.42 Ϯ 4.5 5.4 Ϯ 1.4 -18 Ϯ 2.2 0.0 Ϯ 0.79 V232A ϩ3.6 Ϯ 3.7 5.2 Ϯ 0.4 -18 Ϯ 0.9 ϩ6.1 Ϯ 1.9 P205Scf ϩ4.7 Ϯ 6.0 4.7 Ϯ 1.0 -18 Ϯ 0.7 ϩ4.4 Ϯ 1.6 Q220W ϩ6.3 Ϯ 2.4 5.0 Ϯ 0.7 -21 Ϯ 2.7 -4.9 Ϯ 1.3 G228L ϩ14 Ϯ 5.1 6.9 Ϯ 1.4 -23 Ϯ 1.7 ϩ14 Ϯ 2.6 E217V ϩ28 Ϯ 1.3 6.7 Ϯ 1.0 -25 Ϯ 1.7 ϩ32 Ϯ 4.0 E217F ϩ29 Ϯ 2.8 9.4 Ϯ 1.9 -28 Ϯ 2.6 ϩ17 Ϯ 1.1 E217S/S222E ϩ29 Ϯ 7.6 10 Ϯ 2.3 -25 Ϯ 2.8 ϩ35 Ϯ 0.9 Glycophorin§ - 3.4 Ϯ 0.6 -9.5 Ϯ 1.2 ϩ83 Ϯ 5.1 *Mutant hairpins are listed in order of increasing gel shift. Login to comment
97 PA/VD and ES/SE denote the P205A/V232D and E217S/S222E hairpins, respectively. Login to comment
106 In the presence of the iso-hydropathic mutant E217S/S222E, correlations between each parameter and hydropathy range from R2 values of 0.371 (for bound SDS vs. hydropathy, Fig. 4) to 0.582 (for gel shift vs. hydropathy, Fig. S3). Login to comment
127 It is possible that the 9-10 g SDS/g stoichiometry of E217F and the iso-hydropathic E217S/S222E mutant are representative of detergent loading by a fully ''denatured`` helical membrane protein (such as the ''caterpillar`` structure in Fig. 5F). Login to comment
137 The E217S/S222E mutant is marked with an asterisk. Login to comment
138 Correlation coefficients (R2) of the best fit line in the absence of E217S/S222E (red) or with all mutants (black) are shown. Login to comment
139 Trendline P value in the absence of E217S/S222E is 0.002. Login to comment