4XOU

Crystal structure of the SR Ca2+-ATPase in the Ca2-E1-MgAMPPCP form determined by serial femtosecond crystallography using an X-ray free-electron laser.

Summary for 4XOU

• Entry DOI: 10.2210/pdb4xou/pdb
• Related: 3N8G
• Descriptor: Sarcoplasmic/endoplasmic reticulum calcium ATPase 1, CALCIUM ION, POTASSIUM ION, ... (4 entities in total)
• Functional Keywords: p-type atpase, serial femtosecond crystallography, x-ray free electron laser, hydrolase
• Biological source: Oryctolagus cuniculus (Rabbit)
• Total number of polymer chains: 1
• Total formula weight: 110267.12

Authors

Bublitz, M.,Nass, K.,Drachmann, N.D.,Markvardsen, A.J.,Gutmann, M.J.,Barends, T.R.M.,Mattle, D.,Shoeman, R.L.,Doak, R.B.,Boutet, S.,Messerschmidt, M.,Seibert, M.M.,Williams, G.J.,Foucar, L.,Reinhard, L.,Sitsel, O.,Gregersen, J.L.,Clausen, J.D.,Boesen, T.,Gotfryd, K.,Wang, K.-T.,Olesen, C.,Moller, J.V.,Nissen, P.,Schlichting, I. (deposition date: 2015-01-16, release date: 2015-06-10, Last modification date: 2024-01-10)

Primary citation

Bublitz, M.,Nass, K.,Drachmann, N.D.,Markvardsen, A.J.,Gutmann, M.J.,Barends, T.R.,Mattle, D.,Shoeman, R.L.,Doak, R.B.,Boutet, S.,Messerschmidt, M.,Seibert, M.M.,Williams, G.J.,Foucar, L.,Reinhard, L.,Sitsel, O.,Gregersen, J.L.,Clausen, J.D.,Boesen, T.,Gotfryd, K.,Wang, K.T.,Olesen, C.,Moller, J.V.,Nissen, P.,Schlichting, I.
Structural studies of P-type ATPase-ligand complexes using an X-ray free-electron laser.
Iucrj, 2:409-420, 2015

PubMed 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.

PubMed: 26175901
DOI: 10.1107/S2052252515008969

Experimental method

X-RAY DIFFRACTION (2.8 Å)