Random mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channels
Several inwardly-rectifying (Kir) potassium channels (Kin l 1, Kir41 and Kir4 2) are characterised by their sensitivity to inhibition by intracellular H+ within the physiological range The mechanism by which these channels are regulated by intracellular pH has been the subject of intense scrutiny fo...
| Autores principales: | , , , , |
|---|---|
| Formato: | Journal article |
| Lenguaje: | English |
| Publicado: |
LANDES BIOSCIENCE
2010
|
| _version_ | 1826292372434059264 |
|---|---|
| author | Paynter, J Shang, L Bollepalli, M Baukrowitz, T Tucker, S |
| author_facet | Paynter, J Shang, L Bollepalli, M Baukrowitz, T Tucker, S |
| author_sort | Paynter, J |
| collection | OXFORD |
| description | Several inwardly-rectifying (Kir) potassium channels (Kin l 1, Kir41 and Kir4 2) are characterised by their sensitivity to inhibition by intracellular H+ within the physiological range The mechanism by which these channels are regulated by intracellular pH has been the subject of intense scrutiny for over a decade, yet the molecular identity of the titratable pH-sensor remains elusive In this study we have taken advantage of the acidic intracellular environment of S cerevisiae and used a K+-auxotrophic strain to screen for mutants of Kin 1 1 with impaired pH-sensitivity In addition to the previously identified K80M mutation, this unbiased screening approach identified a novel mutation (S172T) in the second transmembrane domain (TM2) that also produces a marked reduction in pH-sensitivity through destabilization of the closed-state However, despite this extensive mutagenic approach, no mutations could be identified which removed channel pH-sensitivity or which were likely to act as a separate H+-sensor unique to the pH-sensitive Kir channels In order to explain these results we propose a model in which the pH-sensing mechanism is part of an intrinsic gating mechanism common to all Kir channels not just the pH-sensitive Kin channels In this model mutations which disrupt this pH-sensor would result in an increase, not reduction, in pH-sensitivity This has major implications for any future studies of Kir channel pH-sensitivity and explains why formal identification of these pH-sensing residues still represents a major challenge |
| first_indexed | 2024-03-07T03:13:40Z |
| format | Journal article |
| id | oxford-uuid:b511cfe6-9d2b-41d9-be4f-a4d1d6f2e5c5 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:13:40Z |
| publishDate | 2010 |
| publisher | LANDES BIOSCIENCE |
| record_format | dspace |
| spelling | oxford-uuid:b511cfe6-9d2b-41d9-be4f-a4d1d6f2e5c52022-03-27T04:30:37ZRandom mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channelsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:b511cfe6-9d2b-41d9-be4f-a4d1d6f2e5c5EnglishSymplectic Elements at OxfordLANDES BIOSCIENCE2010Paynter, JShang, LBollepalli, MBaukrowitz, TTucker, SSeveral inwardly-rectifying (Kir) potassium channels (Kin l 1, Kir41 and Kir4 2) are characterised by their sensitivity to inhibition by intracellular H+ within the physiological range The mechanism by which these channels are regulated by intracellular pH has been the subject of intense scrutiny for over a decade, yet the molecular identity of the titratable pH-sensor remains elusive In this study we have taken advantage of the acidic intracellular environment of S cerevisiae and used a K+-auxotrophic strain to screen for mutants of Kin 1 1 with impaired pH-sensitivity In addition to the previously identified K80M mutation, this unbiased screening approach identified a novel mutation (S172T) in the second transmembrane domain (TM2) that also produces a marked reduction in pH-sensitivity through destabilization of the closed-state However, despite this extensive mutagenic approach, no mutations could be identified which removed channel pH-sensitivity or which were likely to act as a separate H+-sensor unique to the pH-sensitive Kir channels In order to explain these results we propose a model in which the pH-sensing mechanism is part of an intrinsic gating mechanism common to all Kir channels not just the pH-sensitive Kin channels In this model mutations which disrupt this pH-sensor would result in an increase, not reduction, in pH-sensitivity This has major implications for any future studies of Kir channel pH-sensitivity and explains why formal identification of these pH-sensing residues still represents a major challenge |
| spellingShingle | Paynter, J Shang, L Bollepalli, M Baukrowitz, T Tucker, S Random mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channels |
| title | Random mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channels |
| title_full | Random mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channels |
| title_fullStr | Random mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channels |
| title_full_unstemmed | Random mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channels |
| title_short | Random mutagenesis screening indicates the absence of a separate H+-sensor in the pH-sensitive Kir channels |
| title_sort | random mutagenesis screening indicates the absence of a separate h sensor in the ph sensitive kir channels |
| work_keys_str_mv | AT paynterj randommutagenesisscreeningindicatestheabsenceofaseparatehsensorinthephsensitivekirchannels AT shangl randommutagenesisscreeningindicatestheabsenceofaseparatehsensorinthephsensitivekirchannels AT bollepallim randommutagenesisscreeningindicatestheabsenceofaseparatehsensorinthephsensitivekirchannels AT baukrowitzt randommutagenesisscreeningindicatestheabsenceofaseparatehsensorinthephsensitivekirchannels AT tuckers randommutagenesisscreeningindicatestheabsenceofaseparatehsensorinthephsensitivekirchannels |