ArticleBiophysical journal1997
How to make tubular crystals by reconstitution of detergent-solubilized Ca2(+)-ATPase.
Article in Biophysical journal, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 66 citations in OpenAlex.
- Sarcolipin is a Conformation-Dependent Regulator of the Sarcoplasmic Reticulum Calcium Pump SERCA.The Journal of membrane biology · 2026Article
- Supercharging the calcium pump: Identification of an activation hotspot on SERCA by cryo-EM.bioRxiv : the preprint server for biology · 2026Article
- Stimulation of CaChemMedChem · 2021Article
- Helical Membrane Protein Crystallization in the New Era of Electron Cryo-Microscopy.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Interaction of a Sarcolipin Pentamer and Monomer with the Sarcoplasmic Reticulum Calcium Pump, SERCA.Biophysical journal · 2020Article
- Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Structures of the heart specific SERCA2a CaThe EMBO journal · 2019Article
- The Phospholamban Pentamer Alters Function of the Sarcoplasmic Reticulum Calcium Pump SERCA.Biophysical journal · 2019Article
- ProLIF - quantitative integrin protein-protein interactions and synergistic membrane effects on proteoliposomes.Journal of cell science · 2018Article
- Structure/activity relationship of thapsigargin inhibition on the purified Golgi/secretory pathway CaThe Journal of biological chemistry · 2017Article
- Deducing the symmetry of helical assemblies: Applications to membrane proteins.Journal of structural biology · 2016Article
- Phospholamban C-terminal residues are critical determinants of the structure and function of the calcium ATPase regulatory complex.The Journal of biological chemistry · 2014Article
- Allosteric regulation of SERCA by phosphorylation-mediated conformational shift of phospholamban.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Reconstitution of a Kv channel into lipid membranes for structural and functional studies.Journal of visualized experiments : JoVE · 2013Article
- Lethal, hereditary mutants of phospholamban elude phosphorylation by protein kinase A.The Journal of biological chemistry · 2012Article
- Activating and deactivating roles of lipid bilayers on the Ca(2+)-ATPase/phospholamban complex.Biochemistry · 2011Article
- The architecture of CopA from Archeaoglobus fulgidus studied by cryo-electron microscopy and computational docking.Structure (London, England : 1993) · 2011Article
- An automated pipeline to screen membrane protein 2D crystallization.Journal of structural and functional genomics · 2010Article
- Fourier-Bessel reconstruction of helical assemblies.Methods in enzymology · 2010Article
- Two-dimensional crystallization of integral membrane proteins for electron crystallography.Methods in molecular biology (Clifton, N.J.) · 2010Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
In an attempt to better define the parameters governing reconstitution and two-dimensional crystallization of membrane proteins, we have studied Ca2(+)-ATPase from rabbit sarcoplasmic reticulum. This ion pump forms vanadate-induced crystals in its native membrane and has previously been reconstituted at high lipid-to-protein ratios for functional studies. We have characterized the reconstitution of purified Ca2(+)-ATPase at low lipid-to-protein ratios and discovered procedures that produce long, tubular crystals suitable for helical reconstruction. C12E8 (n-dodecyl-octaethylene-glycol monoether) was used to fully solubilize various mixtures of lipid and purified Ca2(+)-ATPase, and BioBeads were then used to remove the C12E8. Slow removal resulted in two populations of vesicles, and the proteoliposome population was separated from the liposome population on a sucrose density gradient. These proteoliposomes had a lipid-to-protein ratio of 1:2, and virtually 100% of molecules faced the outside of vesicles, as determined by fluorescein isothiocyanate labeling. Cycles of freeze-thaw caused considerable aggregation of these proteoliposomes, and, if phosphatidyl ethanolamine and phosphatidic acid were included, or if the bilayers were doped with small amounts of C12E8, vanadate-induced tubular crystals grew from the aggregates. Thus our procedure comprised two steps-reconstitution followed by crystallization-allowing us to consider mechanisms of bilayer formation separately from those of crystallization and tube formation.
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