Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop
Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topolo...
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E-Artikel |
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Englisch |
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2000 |
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10 |
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Springer Online Journal Archives 1860-2002 |
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in: Pflügers Archiv - 1868, 440(2000) vom: Apr., Seite 556-565 |
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volume:440 ; year:2000 ; month:04 ; pages:556-565 ; extent:10 |
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| 520 | |a Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. | ||
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(DE-627)NLEJ201020718 DE-627 ger DE-627 rakwb eng Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop 2000 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. Springer Online Journal Archives 1860-2002 Becchetti, Andrea oth Roncaglia, Paola oth in Pflügers Archiv 1868 440(2000) vom: Apr., Seite 556-565 (DE-627)NLEJ188987363 (DE-600)1463014-x 1432-2013 nnns volume:440 year:2000 month:04 pages:556-565 extent:10 http://dx.doi.org/10.1007/s004240000324 GBV_USEFLAG_U ZDB-1-SOJ GBV_NL_ARTICLE AR 440 2000 4 556-565 10 |
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(DE-627)NLEJ201020718 DE-627 ger DE-627 rakwb eng Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop 2000 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. Springer Online Journal Archives 1860-2002 Becchetti, Andrea oth Roncaglia, Paola oth in Pflügers Archiv 1868 440(2000) vom: Apr., Seite 556-565 (DE-627)NLEJ188987363 (DE-600)1463014-x 1432-2013 nnns volume:440 year:2000 month:04 pages:556-565 extent:10 http://dx.doi.org/10.1007/s004240000324 GBV_USEFLAG_U ZDB-1-SOJ GBV_NL_ARTICLE AR 440 2000 4 556-565 10 |
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(DE-627)NLEJ201020718 DE-627 ger DE-627 rakwb eng Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop 2000 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. Springer Online Journal Archives 1860-2002 Becchetti, Andrea oth Roncaglia, Paola oth in Pflügers Archiv 1868 440(2000) vom: Apr., Seite 556-565 (DE-627)NLEJ188987363 (DE-600)1463014-x 1432-2013 nnns volume:440 year:2000 month:04 pages:556-565 extent:10 http://dx.doi.org/10.1007/s004240000324 GBV_USEFLAG_U ZDB-1-SOJ GBV_NL_ARTICLE AR 440 2000 4 556-565 10 |
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(DE-627)NLEJ201020718 DE-627 ger DE-627 rakwb eng Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop 2000 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. Springer Online Journal Archives 1860-2002 Becchetti, Andrea oth Roncaglia, Paola oth in Pflügers Archiv 1868 440(2000) vom: Apr., Seite 556-565 (DE-627)NLEJ188987363 (DE-600)1463014-x 1432-2013 nnns volume:440 year:2000 month:04 pages:556-565 extent:10 http://dx.doi.org/10.1007/s004240000324 GBV_USEFLAG_U ZDB-1-SOJ GBV_NL_ARTICLE AR 440 2000 4 556-565 10 |
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(DE-627)NLEJ201020718 DE-627 ger DE-627 rakwb eng Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop 2000 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. Springer Online Journal Archives 1860-2002 Becchetti, Andrea oth Roncaglia, Paola oth in Pflügers Archiv 1868 440(2000) vom: Apr., Seite 556-565 (DE-627)NLEJ188987363 (DE-600)1463014-x 1432-2013 nnns volume:440 year:2000 month:04 pages:556-565 extent:10 http://dx.doi.org/10.1007/s004240000324 GBV_USEFLAG_U ZDB-1-SOJ GBV_NL_ARTICLE AR 440 2000 4 556-565 10 |
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Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop |
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Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. |
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Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. |
| abstract_unstemmed |
Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop. |
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<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">NLEJ201020718</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20210706064807.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">070527s2000 xx |||||o 00| ||eng c</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)NLEJ201020718</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2000</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">10</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zzz</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">z</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zu</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Abstract. In cyclic nucleotide-gated (CNG) channels from the bovine rod, the pore loop "P-loop", connecting the S5 and S6 transmembrane segments, is formed by the residues R345–S371 (here named R1–S27). It determines channel selectivity and contributes to gating. We have studied its topology, by testing the accessibility to Cd2+ of serially substituted cysteine residues. Channels were expressed in Xenopus oocytes. The accessibility of V4C, S6C, T16C, I17C, T20C, P22C and S27C from the cytoplasmic side of the plasma membrane was tested by applying 1–100 µM Cd2+ to the inner face of inside-out patches, at negative membrane potentials. Under these conditions, the effect of Cd2+ on wild-type channels was negligible. The accessibility of the same residues from the external side of the membrane was tested by measuring CNG current inhibition persisting after wash-out of Cd2+ applied to outside-out patches. T16C and I17C channels were strongly inhibited by Cd2+ from the inside, in the presence of cGMP. The K d for T16C block was 16 µM. Thus the T16 and I17 residues participate directly in channel function and are accessible from the cytoplasmic side when the channels are open. In contrast, V4C, T20C and P22C residues were only inhibited when 100 µM Cd2+ was applied externally, suggesting that V4C, T20C and P22C face the outer side of the P-loop.</subfield></datafield><datafield tag="533" ind1=" " ind2=" "><subfield code="f">Springer Online Journal Archives 1860-2002</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Becchetti, Andrea</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Roncaglia, Paola</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">in</subfield><subfield code="t">Pflügers Archiv</subfield><subfield code="d">1868</subfield><subfield code="g">440(2000) vom: Apr., Seite 556-565</subfield><subfield code="w">(DE-627)NLEJ188987363</subfield><subfield code="w">(DE-600)1463014-x</subfield><subfield code="x">1432-2013</subfield><subfield code="7">nnns</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:440</subfield><subfield code="g">year:2000</subfield><subfield code="g">month:04</subfield><subfield code="g">pages:556-565</subfield><subfield code="g">extent:10</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">http://dx.doi.org/10.1007/s004240000324</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ZDB-1-SOJ</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_NL_ARTICLE</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">440</subfield><subfield code="j">2000</subfield><subfield code="c">4</subfield><subfield code="h">556-565</subfield><subfield code="g">10</subfield></datafield></record></collection>
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