ABCC7 p.Gln98Lys
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PMID: 23083715
[PubMed]
El Hiani Y et al: "Tuning of CFTR chloride channel function by location of positive charges within the pore."
No.
Sentence
Comment
49
conductance (Fig. 2, B and C), especially in Q98K (conductance 75 5 1% of WT, n &#bc; 6) and V345K (64 5 3% of WT, n &#bc; 9), and in no case was conductance increased by the addition of a second positive charge.
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ABCC7 p.Gln98Lys 23083715:49:45
status: NEW55 Additional mutations in a K95Q background to transplant the positive charge to pore-lining positions in TM1 (Q98K) or TM6 (I344K, V345K, M348K, and A349K) partially restored NPPB block (Fig. 3), although in no case was the block as strong as for the WT.
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ABCC7 p.Gln98Lys 23083715:55:109
status: NEW56 The rank order of the apparent strength of NPPB block was WT > K95Q/V345K > K95Q/I344K > K95Q/Q98K ~ K95Q/ M348K ~ K95Q/A349K (Fig. 3 B).
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ABCC7 p.Gln98Lys 23083715:56:94
status: NEW60 As shown in Fig. 4, block by Pt(NO2)4 2 was significantly strengthened in each of the mutants Q98K, I344K, V345K, M348K, and A349K, as well as in the previously unstudied S341K.
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ABCC7 p.Gln98Lys 23083715:60:95
status: NEW61 At 0 mV membrane potential, the apparent Kd for Pt(NO2)4 2 block was in the rank order V345K (3.3 5 0.9 mM, n &#bc; 7) % I344K (4.5 5 0.7 mM, n &#bc; 6) < S341K (26.6 5 1.8 mM, n &#bc; 7) < M348K (80.9 5 7.2 mM, n &#bc; 5) % Q98K (95.4 5 11.0 mM, n &#bc; 6) % A349K (117.4 5 7.7 mM, FIGURE 2 Single-channel conductance is restored by moving the positive charge from K95.
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ABCC7 p.Gln98Lys 23083715:61:226
status: NEW74 Blocker voltage dependence was also significantly changed in most mutants, with the effective blocker valence (zd) being significantly increased in M348K and significantly decreased in Q98K, V345K, and A349K (Fig. 4 F).
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ABCC7 p.Gln98Lys 23083715:74:185
status: NEW90 However, although a single positive charge is necessary, the addition of a second positive charge to this region of the pore (as in the point mutants Q98K, I344K, V345K, M348K, and A349K) failed to increase conductance above WT levels (Fig. 2), as previously observed for S1141K (8).
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ABCC7 p.Gln98Lys 23083715:90:150
status: NEW99 In fact, the addition of a second positive charge in Q98K, I344K, V345K, M348K, and A349K led to a small, but significant, decrease in conductance (Fig. 2 C).
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ABCC7 p.Gln98Lys 23083715:99:53
status: NEW105 The weakening of blocker binding seen in K95Q is partially reversed by the second site mutations I344K and V345K, and to a lesser extent Q98K, M348K, and A349K.
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ABCC7 p.Gln98Lys 23083715:105:137
status: NEW146 Again this appears to be a relatively nonsite-specific effect of positive charge, since all mutants studied (Q98K, S341K, I344K, V345K, M348K, and A349K) led to significant increase in apparent affinity of Pt(NO2)4 2 block (Fig. 4), as did S1141K (8).
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ABCC7 p.Gln98Lys 23083715:146:109
status: NEW