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Crystal Structure of the Vanadate-Inhibited Ca2+-ATPase

DOI: 10.1016/j.str.2016.02.018 DOI Help

Authors: Johannes d. Clausen (Aarhus University) , Maike Bublitz (Aarhus University) , Bertrand Arnou (Aarhus University) , Claus Olesen (Aarhus University) , Jens peter Andersen (Aarhus University) , Jesper vuust Møller (Aarhus University) , Poul Nissen (Aarhus University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Structure , VOL 24 , PAGES 617 - 623

State: Published (Approved)
Published: April 2016

Abstract: 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 Ca2+-ATPase with bound vanadate in the absence of Ca2+. Vanadate is bound at the catalytic site as a planar VO3− in complex with water and Mg2+ 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 Ca2+-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 Ca2+-ATPase.

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I24-Microfocus Macromolecular Crystallography

Other Facilities: Deutsches Elektronen-Synchrotron DESY; Swiss Light Source; SOLEIL Synchrotron; European Synchrotron Radiation Facility; MAX Laboratory

Added On: 17/11/2016 13:58

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