Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies

dc.contributor.authorHenao, Fernando
dc.contributor.authorDelavoie, Franck
dc.contributor.authorLacapère, Jean-Jacques
dc.contributor.authorMcIntosh, David B
dc.contributor.authorChampeil, Philippe
dc.date.accessioned2021-10-08T07:20:47Z
dc.date.available2021-10-08T07:20:47Z
dc.date.issued2001
dc.description.abstractThe atomic structure of sarcoplasmic reticulum Ca(2+)-ATPase, in a Ca(2+)-bound conformation, has recently been elucidated (Toyoshima, C., Nakasako, M., Nomura, H. & Ogawa, H. (2000) Nature 405, 647-655). Important steps for further understanding the mechanism of ion pumps will be the atomic structural characterization of different key conformational intermediates of the transport cycle, including phosphorylated intermediates. Following our previous report (Champeil, P., Henao, F., Lacapère, J.-J. & McIntosh, D. B. (2000) J. Biol. Chem. 276, 5795-5803), we show here that it is possible to prepare a phosphorylated form of sarcoplasmic reticulum Ca(2+)-ATPase (labeled with fluorescein isothiocyanate) with a week-long stability both in membranes and in mixed lipid-detergent micelles. We show that this phosphorylated fluorescein isothiocyanate-ATPase can form two-dimensional arrays in membranes, similar to those that were used previously to reconstruct from cryoelectron microscopy images the three-dimensional structure of Ca(2+)-free unphosphorylated ATPase. The results also provide hope that crystals of phosphorylated Ca(2+)-ATPase suitable for x-ray crystallography will be achieved.
dc.identifier.apacitationHenao, F., Delavoie, F., Lacapère, J., McIntosh, D. B., & Champeil, P. (2001). Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies. <i>The Journal of Biological Chemistry</i>, 276(26), 24284 - 24285. http://hdl.handle.net/11427/35005en_ZA
dc.identifier.chicagocitationHenao, Fernando, Franck Delavoie, Jean-Jacques Lacapère, David B McIntosh, and Philippe Champeil "Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies." <i>The Journal of Biological Chemistry</i> 276, 26. (2001): 24284 - 24285. http://hdl.handle.net/11427/35005en_ZA
dc.identifier.citationHenao, F., Delavoie, F., Lacapère, J., McIntosh, D.B. & Champeil, P. 2001. Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies. <i>The Journal of Biological Chemistry.</i> 276(26):24284 - 24285. http://hdl.handle.net/11427/35005en_ZA
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.ris TY - Journal Article AU - Henao, Fernando AU - Delavoie, Franck AU - Lacapère, Jean-Jacques AU - McIntosh, David B AU - Champeil, Philippe AB - The atomic structure of sarcoplasmic reticulum Ca(2+)-ATPase, in a Ca(2+)-bound conformation, has recently been elucidated (Toyoshima, C., Nakasako, M., Nomura, H. & Ogawa, H. (2000) Nature 405, 647-655). Important steps for further understanding the mechanism of ion pumps will be the atomic structural characterization of different key conformational intermediates of the transport cycle, including phosphorylated intermediates. Following our previous report (Champeil, P., Henao, F., Lacapère, J.-J. & McIntosh, D. B. (2000) J. Biol. Chem. 276, 5795-5803), we show here that it is possible to prepare a phosphorylated form of sarcoplasmic reticulum Ca(2+)-ATPase (labeled with fluorescein isothiocyanate) with a week-long stability both in membranes and in mixed lipid-detergent micelles. We show that this phosphorylated fluorescein isothiocyanate-ATPase can form two-dimensional arrays in membranes, similar to those that were used previously to reconstruct from cryoelectron microscopy images the three-dimensional structure of Ca(2+)-free unphosphorylated ATPase. The results also provide hope that crystals of phosphorylated Ca(2+)-ATPase suitable for x-ray crystallography will be achieved. DA - 2001 DB - OpenUCT DP - University of Cape Town IS - 26 J1 - The Journal of Biological Chemistry LK - https://open.uct.ac.za PY - 2001 SM - 0021-9258 SM - 1083-351X T1 - Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies TI - Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies UR - http://hdl.handle.net/11427/35005 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/35005
dc.identifier.vancouvercitationHenao F, Delavoie F, Lacapère J, McIntosh DB, Champeil P. Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies. The Journal of Biological Chemistry. 2001;276(26):24284 - 24285. http://hdl.handle.net/11427/35005.en_ZA
dc.language.isoeng
dc.publisher.departmentDivision of Chemical Pathology
dc.publisher.facultyFaculty of Health Sciences
dc.sourceThe Journal of Biological Chemistry
dc.source.journalissue26
dc.source.journalvolume276
dc.source.pagination24284 - 24285
dc.source.urihttps://dx.doi.org/10.1074/jbc.M103905200
dc.subject.otherAnimals
dc.subject.otherCalcium-Transporting ATPases
dc.subject.otherCrystallization
dc.subject.otherEnzyme Stability
dc.subject.otherFluorescein-5-isothiocyanate
dc.subject.otherFluorescent Dyes
dc.subject.otherKinetics
dc.subject.otherPhosphorylation
dc.subject.otherVanadates
dc.subject.otherFluorescent Dyes
dc.subject.otherVanadates
dc.subject.otherCalcium-Transporting ATPases
dc.subject.otherFluorescein-5-isothiocyanate
dc.titlePhosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies
dc.typeJournal Article
uct.type.publicationResearch
uct.type.resourceJournal Article
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