Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP.
The beta cell KATP channel is an octameric complex of four pore-forming subunits (Kir6.2) and four regulatory subunits (SUR1). A truncated isoform of Kir6.2 (Kir6.2DeltaC26), which expresses independently of SUR1, shows intrinsic ATP sensitivity, suggesting that this subunit is primarily responsible...
| প্রধান লেখক: | , , , |
|---|---|
| বিন্যাস: | Journal article |
| ভাষা: | English |
| প্রকাশিত: |
1998
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| _version_ | 1826263256069570560 |
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| author | Trapp, S Proks, P Tucker, S Ashcroft, F |
| author_facet | Trapp, S Proks, P Tucker, S Ashcroft, F |
| author_sort | Trapp, S |
| collection | OXFORD |
| description | The beta cell KATP channel is an octameric complex of four pore-forming subunits (Kir6.2) and four regulatory subunits (SUR1). A truncated isoform of Kir6.2 (Kir6.2DeltaC26), which expresses independently of SUR1, shows intrinsic ATP sensitivity, suggesting that this subunit is primarily responsible for mediating ATP inhibition. We show here that mutation of C166, which lies at the cytosolic end of the second transmembrane domain, to serine (C166S) increases the open probability of Kir6.2DeltaC26 approximately sevenfold by reducing the time the channel spends in a long closed state. Rundown of channel activity is also decreased. Kir6.2DeltaC26 containing the C166S mutation shows a markedly reduced ATP sensitivity: the Ki is reduced from 175 microM to 2.8 mM. Substitution of threonine, alanine, methionine, or phenylalanine at position C166 also reduced the channel sensitivity to ATP and simultaneously increased the open probability. Thus, ATP does not act as an open channel blocker. The inhibitory effects of tolbutamide are reduced in channels composed of SUR1 and Kir6.2 carrying the C166S mutation. Our results are consistent with the idea that C166 plays a role in the intrinsic gating of the channel, possibly by influencing a gate located at the intracellular end of the pore. Kinetic analysis suggests that the apparent decrease in ATP sensitivity, and the changes in other properties, observed when C166 is mutated is largely a consequence of the impaired transition from the open to the long closed state. |
| first_indexed | 2024-03-06T19:48:50Z |
| format | Journal article |
| id | oxford-uuid:2344594f-9a08-4a99-b44e-439bcc0355d9 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:48:50Z |
| publishDate | 1998 |
| record_format | dspace |
| spelling | oxford-uuid:2344594f-9a08-4a99-b44e-439bcc0355d92022-03-26T11:43:27ZMolecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:2344594f-9a08-4a99-b44e-439bcc0355d9EnglishSymplectic Elements at Oxford1998Trapp, SProks, PTucker, SAshcroft, FThe beta cell KATP channel is an octameric complex of four pore-forming subunits (Kir6.2) and four regulatory subunits (SUR1). A truncated isoform of Kir6.2 (Kir6.2DeltaC26), which expresses independently of SUR1, shows intrinsic ATP sensitivity, suggesting that this subunit is primarily responsible for mediating ATP inhibition. We show here that mutation of C166, which lies at the cytosolic end of the second transmembrane domain, to serine (C166S) increases the open probability of Kir6.2DeltaC26 approximately sevenfold by reducing the time the channel spends in a long closed state. Rundown of channel activity is also decreased. Kir6.2DeltaC26 containing the C166S mutation shows a markedly reduced ATP sensitivity: the Ki is reduced from 175 microM to 2.8 mM. Substitution of threonine, alanine, methionine, or phenylalanine at position C166 also reduced the channel sensitivity to ATP and simultaneously increased the open probability. Thus, ATP does not act as an open channel blocker. The inhibitory effects of tolbutamide are reduced in channels composed of SUR1 and Kir6.2 carrying the C166S mutation. Our results are consistent with the idea that C166 plays a role in the intrinsic gating of the channel, possibly by influencing a gate located at the intracellular end of the pore. Kinetic analysis suggests that the apparent decrease in ATP sensitivity, and the changes in other properties, observed when C166 is mutated is largely a consequence of the impaired transition from the open to the long closed state. |
| spellingShingle | Trapp, S Proks, P Tucker, S Ashcroft, F Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP. |
| title | Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP. |
| title_full | Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP. |
| title_fullStr | Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP. |
| title_full_unstemmed | Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP. |
| title_short | Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP. |
| title_sort | molecular analysis of atp sensitive k channel gating and implications for channel inhibition by atp |
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