Crystal structure of the vanadate-inhibited Ca(2+)-ATPase
Vanadate is the hallmark inhibitor of the P-type ATPase family; however, structural details of its inhibitory mechanism have remained unresolved. We have determined the crystal structure of sarcoplasmic reticulum Ca(2+)-ATPase with bound vanadate in the absence of Ca(2+). Vanadate is bound at the ca...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Elsevier
2016
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| _version_ | 1826287769945636864 |
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| author | Clausen, J Bublitz, M Arnou, B Olesen, C Andersen, J Møller, J Nissen, P |
| author_facet | Clausen, J Bublitz, M Arnou, B Olesen, C Andersen, J Møller, J Nissen, P |
| author_sort | Clausen, J |
| collection | OXFORD |
| description | Vanadate is the hallmark inhibitor of the P-type ATPase family; however, structural details of its inhibitory mechanism have remained unresolved. We have determined the crystal structure of sarcoplasmic reticulum Ca(2+)-ATPase with bound vanadate in the absence of Ca(2+). Vanadate is bound at the catalytic site as a planar VO3(-) in complex with water and Mg(2+) in a dephosphorylation transition-state-like conformation. Validating bound VO3(-) by anomalous difference Fourier maps using long-wavelength data we also identify a hitherto undescribed Cl(-) site near the dephosphorylation site. Crystallization was facilitated by trinitrophenyl (TNP)-derivatized nucleotides that bind with the TNP moiety occupying the binding pocket that normally accommodates the adenine of ATP, rationalizing their remarkably high affinity for E2P-like conformations of the Ca(2+)-ATPase. A comparison of the configurations of bound nucleotide analogs in the E2·VO3(-) structure with that in E2·BeF3(-) (E2P ground state analog) reveals multiple binding modes to the Ca(2+)-ATPase. |
| first_indexed | 2024-03-07T02:03:37Z |
| format | Journal article |
| id | oxford-uuid:9e389297-7d92-47f5-9731-f9a506c1b79b |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:03:37Z |
| publishDate | 2016 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:9e389297-7d92-47f5-9731-f9a506c1b79b2022-03-27T00:48:32ZCrystal structure of the vanadate-inhibited Ca(2+)-ATPaseJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:9e389297-7d92-47f5-9731-f9a506c1b79bEnglishSymplectic Elements at OxfordElsevier2016Clausen, JBublitz, MArnou, BOlesen, CAndersen, JMøller, JNissen, PVanadate is the hallmark inhibitor of the P-type ATPase family; however, structural details of its inhibitory mechanism have remained unresolved. We have determined the crystal structure of sarcoplasmic reticulum Ca(2+)-ATPase with bound vanadate in the absence of Ca(2+). Vanadate is bound at the catalytic site as a planar VO3(-) in complex with water and Mg(2+) in a dephosphorylation transition-state-like conformation. Validating bound VO3(-) by anomalous difference Fourier maps using long-wavelength data we also identify a hitherto undescribed Cl(-) site near the dephosphorylation site. Crystallization was facilitated by trinitrophenyl (TNP)-derivatized nucleotides that bind with the TNP moiety occupying the binding pocket that normally accommodates the adenine of ATP, rationalizing their remarkably high affinity for E2P-like conformations of the Ca(2+)-ATPase. A comparison of the configurations of bound nucleotide analogs in the E2·VO3(-) structure with that in E2·BeF3(-) (E2P ground state analog) reveals multiple binding modes to the Ca(2+)-ATPase. |
| spellingShingle | Clausen, J Bublitz, M Arnou, B Olesen, C Andersen, J Møller, J Nissen, P Crystal structure of the vanadate-inhibited Ca(2+)-ATPase |
| title | Crystal structure of the vanadate-inhibited Ca(2+)-ATPase |
| title_full | Crystal structure of the vanadate-inhibited Ca(2+)-ATPase |
| title_fullStr | Crystal structure of the vanadate-inhibited Ca(2+)-ATPase |
| title_full_unstemmed | Crystal structure of the vanadate-inhibited Ca(2+)-ATPase |
| title_short | Crystal structure of the vanadate-inhibited Ca(2+)-ATPase |
| title_sort | crystal structure of the vanadate inhibited ca 2 atpase |
| work_keys_str_mv | AT clausenj crystalstructureofthevanadateinhibitedca2atpase AT bublitzm crystalstructureofthevanadateinhibitedca2atpase AT arnoub crystalstructureofthevanadateinhibitedca2atpase AT olesenc crystalstructureofthevanadateinhibitedca2atpase AT andersenj crystalstructureofthevanadateinhibitedca2atpase AT møllerj crystalstructureofthevanadateinhibitedca2atpase AT nissenp crystalstructureofthevanadateinhibitedca2atpase |