Inhibited KdpFABC transitions into an E1 off-cycle state
KdpFABC is a high-affinity prokaryotic K+ uptake system that forms a functional chimera between a channel-like subunit (KdpA) and a P-type ATPase (KdpB). At high K+ levels, KdpFABC needs to be inhibited to prevent excessive K+ accumulation to the point of toxicity. This is achieved by a phosphorylat...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-10-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/80988 |
| _version_ | 1818470928319774720 |
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| author | Jakob M Silberberg Charlott Stock Lisa Hielkema Robin A Corey Jan Rheinberger Dorith Wunnicke Victor RA Dubach Phillip J Stansfeld Inga Hänelt Cristina Paulino |
| author_facet | Jakob M Silberberg Charlott Stock Lisa Hielkema Robin A Corey Jan Rheinberger Dorith Wunnicke Victor RA Dubach Phillip J Stansfeld Inga Hänelt Cristina Paulino |
| author_sort | Jakob M Silberberg |
| collection | DOAJ |
| description | KdpFABC is a high-affinity prokaryotic K+ uptake system that forms a functional chimera between a channel-like subunit (KdpA) and a P-type ATPase (KdpB). At high K+ levels, KdpFABC needs to be inhibited to prevent excessive K+ accumulation to the point of toxicity. This is achieved by a phosphorylation of the serine residue in the TGES162 motif in the A domain of the pump subunit KdpB (KdpBS162-P). Here, we explore the structural basis of inhibition by KdpBS162 phosphorylation by determining the conformational landscape of KdpFABC under inhibiting and non-inhibiting conditions. Under turnover conditions, we identified a new inhibited KdpFABC state that we termed E1P tight, which is not part of the canonical Post-Albers transport cycle of P-type ATPases. It likely represents the biochemically described stalled E1P state adopted by KdpFABC upon KdpBS162 phosphorylation. The E1P tight state exhibits a compact fold of the three cytoplasmic domains and is likely adopted when the transition from high-energy E1P states to E2P states is unsuccessful. This study represents a structural characterization of a biologically relevant off-cycle state in the P-type ATPase family and supports the emerging discussion of P-type ATPase regulation by such states. |
| first_indexed | 2024-04-13T21:44:06Z |
| format | Article |
| id | doaj.art-18dbd7d1afa04058bd4eae055360e3c2 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-13T21:44:06Z |
| publishDate | 2022-10-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-18dbd7d1afa04058bd4eae055360e3c22022-12-22T02:28:39ZengeLife Sciences Publications LtdeLife2050-084X2022-10-011110.7554/eLife.80988Inhibited KdpFABC transitions into an E1 off-cycle stateJakob M Silberberg0https://orcid.org/0000-0003-1721-8666Charlott Stock1https://orcid.org/0000-0001-5471-3696Lisa Hielkema2Robin A Corey3https://orcid.org/0000-0003-1820-7993Jan Rheinberger4https://orcid.org/0000-0002-9901-2065Dorith Wunnicke5Victor RA Dubach6https://orcid.org/0000-0002-1657-7184Phillip J Stansfeld7Inga Hänelt8https://orcid.org/0000-0003-1495-3163Cristina Paulino9https://orcid.org/0000-0001-7017-109XInstitute of Biochemistry, Biocenter, Goethe University Frankfurt, Frankfurt, GermanyInstitute of Biochemistry, Biocenter, Goethe University Frankfurt, Frankfurt, GermanyDepartment of Structural Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, NetherlandsDepartment of Biochemistry, University of Oxford, Oxford, United KingdomDepartment of Structural Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, NetherlandsInstitute of Biochemistry, Biocenter, Goethe University Frankfurt, Frankfurt, GermanyDepartment of Structural Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, NetherlandsSchool of Life Sciences & Department of Chemistry, University of Warwick, Coventry, United KingdomInstitute of Biochemistry, Biocenter, Goethe University Frankfurt, Frankfurt, GermanyDepartment of Structural Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, NetherlandsKdpFABC is a high-affinity prokaryotic K+ uptake system that forms a functional chimera between a channel-like subunit (KdpA) and a P-type ATPase (KdpB). At high K+ levels, KdpFABC needs to be inhibited to prevent excessive K+ accumulation to the point of toxicity. This is achieved by a phosphorylation of the serine residue in the TGES162 motif in the A domain of the pump subunit KdpB (KdpBS162-P). Here, we explore the structural basis of inhibition by KdpBS162 phosphorylation by determining the conformational landscape of KdpFABC under inhibiting and non-inhibiting conditions. Under turnover conditions, we identified a new inhibited KdpFABC state that we termed E1P tight, which is not part of the canonical Post-Albers transport cycle of P-type ATPases. It likely represents the biochemically described stalled E1P state adopted by KdpFABC upon KdpBS162 phosphorylation. The E1P tight state exhibits a compact fold of the three cytoplasmic domains and is likely adopted when the transition from high-energy E1P states to E2P states is unsuccessful. This study represents a structural characterization of a biologically relevant off-cycle state in the P-type ATPase family and supports the emerging discussion of P-type ATPase regulation by such states.https://elifesciences.org/articles/80988cryo-EMP-type ATPasemembrane transport mechanismprotein regulationpotassium uptakestructure–function |
| spellingShingle | Jakob M Silberberg Charlott Stock Lisa Hielkema Robin A Corey Jan Rheinberger Dorith Wunnicke Victor RA Dubach Phillip J Stansfeld Inga Hänelt Cristina Paulino Inhibited KdpFABC transitions into an E1 off-cycle state eLife cryo-EM P-type ATPase membrane transport mechanism protein regulation potassium uptake structure–function |
| title | Inhibited KdpFABC transitions into an E1 off-cycle state |
| title_full | Inhibited KdpFABC transitions into an E1 off-cycle state |
| title_fullStr | Inhibited KdpFABC transitions into an E1 off-cycle state |
| title_full_unstemmed | Inhibited KdpFABC transitions into an E1 off-cycle state |
| title_short | Inhibited KdpFABC transitions into an E1 off-cycle state |
| title_sort | inhibited kdpfabc transitions into an e1 off cycle state |
| topic | cryo-EM P-type ATPase membrane transport mechanism protein regulation potassium uptake structure–function |
| url | https://elifesciences.org/articles/80988 |
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