ABCC7 p.Gln220Gly

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PMID: 17516627 [PubMed] Wehbi H et al: "Role of the extracellular loop in the folding of a CFTR transmembrane helical hairpin."
No. Sentence Comment
66 We assessed the role of Q220 in TM3/4 folding by constructing two point mutants that could have variable effects on side chain electrostatic interactions: Q220G and Q220N.
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ABCC7 p.Gln220Gly 17516627:66:155
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67 However, neither removal of the polar side chain in the Q220G mutant nor truncation of side chain length by a methylene group in the Q220N mutant effected a significant change in hairpin mobility (Figure 3, Table 1), suggesting that the WT Q220 was not likely to be participating in ECL2 side chain-side chain interactions.
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ABCC7 p.Gln220Gly 17516627:67:56
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83 The Q220E replacement migrates faster than TM3/4 WT, the Q220G and Q220N mutants migrate at equivalent rates to TM3/4 WT, while the Q220W, Q220K, and Q220R substitutions migrate more slowly.
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ABCC7 p.Gln220Gly 17516627:83:57
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87 While Q220G- and Q220N-substituted hairpins migrate at the same rate as WT TM3/4, we undertook to examine this position systematically to determine the molecular determinants of migration rate variation.
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ABCC7 p.Gln220Gly 17516627:87:6
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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.
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ABCC7 p.Gln220Gly 17516627:97:545
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