Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States

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dc.contributor.authorClausen, Johannes D
dc.contributor.authorMcIntosh, David B
dc.contributor.authorWoolley, David G
dc.contributor.authorAnthonisen, Anne Nyholm
dc.contributor.authorVilsen, Bente
dc.contributor.authorAndersen, Jens Peter
dc.date.accessioned2021-10-08T07:22:51Z
dc.date.available2021-10-08T07:22:51Z
dc.date.issued2006
dc.description.abstractMutants with alteration to Asn(706) of the highly conserved (701)TGDGVND(707) motif in domain P of sarcoplasmic reticulum Ca(2+)-ATPase were analyzed for changes in transport cycle kinetics and binding of the inhibitors vanadate, BeF, AlF, and MgF. The fluorides likely mimic the phosphoryl group/P(i) in the respective ground, transition, and product states of phosphoenzyme hydrolysis (Danko, S., Yamasaki, K., Daiho, T., and Suzuki, H. (2004) J. Biol. Chem. 279, 14991-14998). Binding of BeF, AlF, and MgF was also studied for mutant Glu(183) --> Ala, where the glutamate of the (181)TGES(184) motif in domain A is replaced. Mutations of Asn(706) and Glu(183) have in common that they dramatically impede the function of the enzyme in E2 states, but have little effect in E1. Contrary to the Glu(183) mutant, in which E2P slowly accumulates (Clausen, J. D., Vilsen, B., McIntosh, D. B., Einholm, A. P., and Andersen, J. P. (2004) Proc. Natl. Acad. Sci. U. S. A. 101, 2776-2781), E2P formation was not detectable with the Asn(706) mutants. Differential sensitivities of the mutants to inhibition by AlF, MgF, and BeF made it possible to distinguish different roles of Asn(706) and Glu(183). Hence, Asn(706) is less important than Glu(183) for gaining the transition state during E2P hydrolysis but plays critical roles in stabilization of E2P ground and E2.P(i) product states and in the major conformational changes associated with the Ca(2)E1P --> E2P and E2 --> Ca(2)E1 transitions, which seem to be facilitated by interaction of Asn(706) with domain A.
dc.identifier.apacitationClausen, J. D., McIntosh, D. B., Woolley, D. G., Anthonisen, A. N., Vilsen, B., & Andersen, J. P. (2006). Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States. <i>The Journal of Biological Chemistry</i>, 281(14), 9471 - 9481. http://hdl.handle.net/11427/35011en_ZA
dc.identifier.chicagocitationClausen, Johannes D, David B McIntosh, David G Woolley, Anne Nyholm Anthonisen, Bente Vilsen, and Jens Peter Andersen "Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States." <i>The Journal of Biological Chemistry</i> 281, 14. (2006): 9471 - 9481. http://hdl.handle.net/11427/35011en_ZA
dc.identifier.citationClausen, J.D., McIntosh, D.B., Woolley, D.G., Anthonisen, A.N., Vilsen, B. & Andersen, J.P. 2006. Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States. <i>The Journal of Biological Chemistry.</i> 281(14):9471 - 9481. http://hdl.handle.net/11427/35011en_ZA
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.ris TY - Journal Article AU - Clausen, Johannes D AU - McIntosh, David B AU - Woolley, David G AU - Anthonisen, Anne Nyholm AU - Vilsen, Bente AU - Andersen, Jens Peter AB - Mutants with alteration to Asn(706) of the highly conserved (701)TGDGVND(707) motif in domain P of sarcoplasmic reticulum Ca(2+)-ATPase were analyzed for changes in transport cycle kinetics and binding of the inhibitors vanadate, BeF, AlF, and MgF. The fluorides likely mimic the phosphoryl group/P(i) in the respective ground, transition, and product states of phosphoenzyme hydrolysis (Danko, S., Yamasaki, K., Daiho, T., and Suzuki, H. (2004) J. Biol. Chem. 279, 14991-14998). Binding of BeF, AlF, and MgF was also studied for mutant Glu(183) --> Ala, where the glutamate of the (181)TGES(184) motif in domain A is replaced. Mutations of Asn(706) and Glu(183) have in common that they dramatically impede the function of the enzyme in E2 states, but have little effect in E1. Contrary to the Glu(183) mutant, in which E2P slowly accumulates (Clausen, J. D., Vilsen, B., McIntosh, D. B., Einholm, A. P., and Andersen, J. P. (2004) Proc. Natl. Acad. Sci. U. S. A. 101, 2776-2781), E2P formation was not detectable with the Asn(706) mutants. Differential sensitivities of the mutants to inhibition by AlF, MgF, and BeF made it possible to distinguish different roles of Asn(706) and Glu(183). Hence, Asn(706) is less important than Glu(183) for gaining the transition state during E2P hydrolysis but plays critical roles in stabilization of E2P ground and E2.P(i) product states and in the major conformational changes associated with the Ca(2)E1P --> E2P and E2 --> Ca(2)E1 transitions, which seem to be facilitated by interaction of Asn(706) with domain A. DA - 2006 DB - OpenUCT DP - University of Cape Town IS - 14 J1 - The Journal of Biological Chemistry LK - https://open.uct.ac.za PY - 2006 SM - 0021-9258 SM - 1083-351X T1 - Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States TI - Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States UR - http://hdl.handle.net/11427/35011 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/35011
dc.identifier.vancouvercitationClausen JD, McIntosh DB, Woolley DG, Anthonisen AN, Vilsen B, Andersen JP. Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States. The Journal of Biological Chemistry. 2006;281(14):9471 - 9481. http://hdl.handle.net/11427/35011.en_ZA
dc.language.isoeng
dc.publisher.departmentInstitute of Infectious Disease and Molecular Medicine
dc.publisher.facultyFaculty of Health Sciences
dc.sourceThe Journal of Biological Chemistry
dc.source.journalissue14
dc.source.journalvolume281
dc.source.pagination9471 - 9481
dc.source.urihttps://dx.doi.org/10.1074/jbc.M512371200
dc.subject.otherAdenosine Triphosphate
dc.subject.otherAnimals
dc.subject.otherAsparagine
dc.subject.otherBinding Sites
dc.subject.otherCalcium
dc.subject.otherCalcium-Transporting ATPases
dc.subject.otherCatalytic Domain
dc.subject.otherEnzyme Activation
dc.subject.otherEnzyme Inhibitors
dc.subject.otherFluorides
dc.subject.otherGlutamic Acid
dc.subject.otherHydrolysis
dc.subject.otherKinetics
dc.subject.otherMutagenesis, Site-Directed
dc.subject.otherPhosphates
dc.subject.otherPoint Mutation
dc.subject.otherProtein Binding
dc.subject.otherProtein Conformation
dc.titleAsparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States
dc.typeJournal Article
uct.type.publicationResearch
uct.type.resourceJournal Article
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