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

Maike Bublitz, Karol Nass, Nikolaj D. Drachmann, Anders J. Markvardsen, Matthias J. Gutmann, Thomas R.M. Barends, Daniel Mattle, Robert L. Shoeman, R. Bruce Doak, Sébastien Boutet, Marc Messerschmidt, Marvin M. Seibert, Garth J. Williams, Lutz Foucar, Linda Reinhard, Oleg Sitsel, Jonas L. Gregersen, Johannes D. Clausen, Thomas Boesen, Kamil Gotfryd & 5 others Kai Tuo Wang, Claus Olesen, Jesper V. Møller, Poul Nissen, Ilme Schlichting

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)409-420
Number of pages12
JournalIUCrJ
Volume2
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

Fingerprint

X ray lasers
Free electron lasers
free electron lasers
Adenosine Triphosphatases
Lasers
Ligands
X-Rays
Electrons
ligands
proteins
Proteins
Membrane Proteins
x rays
membranes
Membranes
inhibitors
Crystallography
Microcrystals
Vanadates
metabolites

Keywords

  • ligand screening
  • P-type ATPases
  • serial femtosecond crystallography
  • XFEL

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics

Cite this

Bublitz, M., Nass, K., Drachmann, N. D., Markvardsen, A. J., Gutmann, M. J., Barends, T. R. M., ... Schlichting, I. (2015). Structural studies of P-type ATPase-ligand complexes using an X-ray free-electron laser. IUCrJ, 2, 409-420. https://doi.org/10.1107/S2052252515008969

Structural studies of P-type ATPase-ligand complexes using an X-ray free-electron laser. / Bublitz, Maike; Nass, Karol; Drachmann, Nikolaj D.; Markvardsen, Anders J.; Gutmann, Matthias J.; Barends, Thomas R.M.; Mattle, Daniel; Shoeman, Robert L.; Doak, R. Bruce; Boutet, Sébastien; Messerschmidt, Marc; Seibert, Marvin M.; Williams, Garth J.; Foucar, Lutz; Reinhard, Linda; Sitsel, Oleg; Gregersen, Jonas L.; Clausen, Johannes D.; Boesen, Thomas; Gotfryd, Kamil; Wang, Kai Tuo; Olesen, Claus; Møller, Jesper V.; Nissen, Poul; Schlichting, Ilme.

In: IUCrJ, Vol. 2, 01.01.2015, p. 409-420.

Research output: Contribution to journalArticle

Bublitz, M, Nass, K, Drachmann, ND, Markvardsen, AJ, Gutmann, MJ, Barends, TRM, Mattle, D, Shoeman, RL, Doak, RB, Boutet, S, Messerschmidt, M, Seibert, MM, Williams, GJ, Foucar, L, Reinhard, L, Sitsel, O, Gregersen, JL, Clausen, JD, Boesen, T, Gotfryd, K, Wang, KT, Olesen, C, Møller, JV, Nissen, P & Schlichting, I 2015, 'Structural studies of P-type ATPase-ligand complexes using an X-ray free-electron laser', IUCrJ, vol. 2, pp. 409-420. https://doi.org/10.1107/S2052252515008969
Bublitz M, Nass K, Drachmann ND, Markvardsen AJ, Gutmann MJ, Barends TRM et al. Structural studies of P-type ATPase-ligand complexes using an X-ray free-electron laser. IUCrJ. 2015 Jan 1;2:409-420. https://doi.org/10.1107/S2052252515008969
Bublitz, Maike ; Nass, Karol ; Drachmann, Nikolaj D. ; Markvardsen, Anders J. ; Gutmann, Matthias J. ; Barends, Thomas R.M. ; Mattle, Daniel ; Shoeman, Robert L. ; Doak, R. Bruce ; Boutet, Sébastien ; Messerschmidt, Marc ; Seibert, Marvin M. ; Williams, Garth J. ; Foucar, Lutz ; Reinhard, Linda ; Sitsel, Oleg ; Gregersen, Jonas L. ; Clausen, Johannes D. ; Boesen, Thomas ; Gotfryd, Kamil ; Wang, Kai Tuo ; Olesen, Claus ; Møller, Jesper V. ; Nissen, Poul ; Schlichting, Ilme. / Structural studies of P-type ATPase-ligand complexes using an X-ray free-electron laser. In: IUCrJ. 2015 ; Vol. 2. pp. 409-420.
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