Structural studies of P-type ATPase-ligand complexes using an X-ray free-electron laser

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

DOI

  • Maike Bublitz
  • ,
  • Karol Nass
  • ,
  • Nikolaj D Drachmann
  • ,
  • Anders J Markvardsen, Danmark
  • Matthias J Gutmann, Danmark
  • Thomas R M Barends
  • ,
  • Daniel Mattle
  • ,
  • Robert L Shoeman
  • ,
  • R Bruce Doak
  • ,
  • Sébastien Boutet
  • ,
  • Marc Messerschmidt
  • ,
  • Marvin M Seibert, Danmark
  • Garth J Williams
  • ,
  • Lutz Foucar
  • ,
  • Linda Reinhard
  • ,
  • Oleg Sitsel
  • ,
  • Jonas L Gregersen
  • ,
  • Johannes D Clausen
  • ,
  • Thomas Boesen
  • Kamil Gotfryd
  • ,
  • Kai-Tuo Wang
  • ,
  • Claus Olesen
  • Jesper V Møller
  • Poul Nissen
  • Ilme Schlichting

Membrane proteins are key players in biological systems, mediating signalling events and the specific transport of e.g. ions and metabolites. Consequently, membrane proteins are targeted by a large number of currently approved drugs. Understanding their functions and molecular mechanisms is greatly dependent on structural information, not least on complexes with functionally or medically important ligands. Structure determination, however, is hampered by the difficulty of obtaining well diffracting, macroscopic crystals. Here, the feasibility of X-ray free-electron-laser-based serial femtosecond crystallography (SFX) for the structure determination of membrane protein-ligand complexes using microcrystals of various native-source and recombinant P-type ATPase complexes is demonstrated. The data reveal the binding sites of a variety of ligands, including lipids and inhibitors such as the hallmark P-type ATPase inhibitor orthovanadate. By analyzing the resolution dependence of ligand densities and overall model qualities, SFX data quality metrics as well as suitable refinement procedures are discussed. Even at relatively low resolution and multiplicity, the identification of ligands can be demonstrated. This makes SFX a useful tool for ligand screening and thus for unravelling the molecular mechanisms of biologically active proteins.

OriginalsprogEngelsk
TidsskriftIUCrJ
Vol/bind2
NummerPt 4
Sider (fra-til)409-420
Antal sider12
ISSN2052-2525
DOI
StatusUdgivet - 1 jul. 2015

Se relationer på Aarhus Universitet Citationsformater

ID: 90280772