Quasi-irreversible inactivation of the sarcoplasmic reticulum Ca2+-ATPase by simultaneous tight binding of magnesium and fluoride to the catalytic site

Kubota, Tatsuya; Daiho, Takashi; Kanazawa, Tohru

doi:10.1016/0167-4838(93)90174-p

Cited by 31 publications

(55 citation statements)

References 44 publications

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“…3, B and D). In the case of wild type, the ability to form EP is restored, consistent with the previous observation (25,36) ATP-induced phosphorylation. Therefore, the complex produced in the mutant with BeF x is likely E2Ca 2 ⅐BeF 3 Ϫ , an analog of E2PCa 2 (as is in fact shown later in the Ca 2ϩ binding and structural analyses in Fig.…”

Section: Methodssupporting

confidence: 79%

Stable Structural Analog of Ca2+-ATPase ADP-insensitive Phosphoenzyme with Occluded Ca2+ Formed by Elongation of A-domain/M1′-linker and Beryllium Fluoride Binding

Daiho

Dankó

Yamasaki

et al. 2010

Journal of Biological Chemistry

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During EP isomerization/Ca 2ϩ -release (E1PCa 2 3 E2P ϩ 2Ca 2ϩ ), the A domain swings around parallel to the membrane plane (i.e. horizontal), whereas the A and P domains and M2 incline and tightly associate (Fig. 2) (15-25). We found that shortening of the A/M1Ј-linker by deletion of any single residue blocks E1PCa 2 3 E2PCa 2 isomerization and E2P hydrolysis (26). On the other hand, its elongation by two or more glycine insertions markedly accelerates the isomerization and blocks Ca 2ϩ deocclusion/release (E2PCa 2 3 E2P ϩ 2Ca 2ϩ ) (14). Thus, elongating the A/M1Ј-linker stabilized the normally transient intermediate state E2PCa 2 (i.e. ADP-insensitive EP with occluded Ca 2ϩ ) and showed that the length of this linker is critical for the structural changes that occur during E1PCa 2 3 E2PCa 2 3 E2P ϩ 2Ca 2ϩ and subsequent E2P hydrolysis (14, 26).We have recently developed an E1Ca 2 ⅐BeF 3 Ϫ complex as a stable analog of E1PCa 2 ⅐Mg 2ϩ (E1PCa 2 with bound Mg 2ϩ at the catalytic site) (27). Structural analysis of the analog and intermediate states suggests that formation of native E1PCa 2 ⅐Mg 2ϩ results in structural changes in the cytoplasmic and transmembrane domains due to configuration and ligation changes of the phosphate moiety (27). The Mg 2ϩ bound at the catalytic site contributes to these structural changes (27)

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Section: Methodssupporting

confidence: 79%

Stable Structural Analog of Ca2+-ATPase ADP-insensitive Phosphoenzyme with Occluded Ca2+ Formed by Elongation of A-domain/M1′-linker and Beryllium Fluoride Binding

Daiho

Dankó

Yamasaki

et al. 2010

Journal of Biological Chemistry

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“…The binding affinity observed under these conditions (Fig 8) was considerably lower (37), and determination of binding at high ATP concentrations was prevented by the unfavorable noise to signal ratio. Nevertheless, it is clear that ATP can still bind to the phosphoenzyme intermediate analog, although with low affinity.…”

Section: Atp Binding To the Fluoroaluminate Analog Of Epmentioning

confidence: 90%

“…Recombinant Ca 2+ ATPase was obtained from COS-1 cells infected with adenovirus vectors carrying chicken WT (33) or mutant cDNA (34). ATP and ADP binding was measured by a filtration method (35)(36)(37) For each experimental sample, 3 ml of ice cold medium were added by fast mixing to 0.3 ml medium containing 0.4 mg SR protein, and filtered after 10 seconds through a 0.65 μm Millipore filter. Controls were obtained by passing through the filters the same volume of reaction medium containing corresponding concentrations of nucleotide in the absence of SR protein.…”

Section: Methodsmentioning

confidence: 99%

Concerted Conformational Effects of Ca²⁺and ATP Are Required for Activation of Sequential Reactions in the Ca²⁺ATPase (SERCA) Catalytic Cycle^,

Inesi

Lewis

et al. 2006

Biochemistry

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We relate solution behavior to the crystal structure of the Ca 2+ ATPase (SERCA). We find that nucleotide binding occurs with high affinity through interaction of the adenosine moiety with the N domain, even in the absence of Ca 2+ and Mg 2+ , or to the closed conformation stabilized by thapsigargin (TG). Why then is Ca 2+ crucial for ATP utilization? The influence of AMPPCP, Ca 2+ and Mg 2+ on proteolytic digestion patterns, interpreted in the light of known crystal structures, indicates that a Ca 2+ dependent conformation of the ATPase headpiece is required for a further transition induced by nucleotide binding. This includes opening of the headpiece, which in turn allows inclination of the "A" domain and bending of the "P" domain. Thereby, the phosphate chain of bound ATP acquires an extended configuration allowing the γ-phosphate to reach Asp351 to form a complex including Mg 2+ . We demonstrate by Asp351 mutation that this "productive" conformation of the substrate-enzyme complex is unstable due to electrostatic repulsion at the phosphorylation site. However, this conformation is subsequently stabilized by covalent engagement of the γ-phosphate yielding the phosphoenzyme intermediate. We also demonstrate that the ADP product remains bound with high affinity to the transition state complex, but dissociates with lower affinity as the phosphoenzyme undergoes a further conformational change (i.e., E1P to E2-P transition). Finally, we measured low affinity ATP binding to stable phosphoenzyme analogs, demonstrating that the E1-P to E2-P transition and the enzyme turnover are accelerated by ATP binding to the phosphoenzyme in exchange for ADP.The Ca 2+ ATPase of Sarco-Endoplasmic Reticulum (SERCA) is a membrane bound enzyme that uses ATP as an energy source for Ca 2+ transport. The 100 kD ATPase protein includes ten helical segments partitioned within the membrane bilayer, and a headpiece protruding from the cytosolic membrane surface (1,2). Binding of 2 Ca 2+ is required for enzyme (E) activation, whereby the ATP terminal phosphate is utilized to form a phosphorylated enzyme intermediate (E-P). The bound Ca 2+ then undergoes occlusion and vectorial translocation. The catalytic cycle is completed by hydrolytic cleavage of E-P (3-5)

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“…Free Ca 2ϩ concentrations were calculated as described previously (34). Data were analyzed using Origin software (Microcal Software, Inc., Northampton, MA).…”

Section: Methodsmentioning

confidence: 99%

Comprehensive Analysis of Expression and Function of 51 Sarco(endo)plasmic Reticulum Ca2+-ATPase Mutants Associated with Darier Disease

Miyauchi

Daiho

Yamasaki

et al. 2006

Journal of Biological Chemistry

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We examined possible defects of sarco(endo)plasmic reticulum Ca catalyze Ca 2ϩ transport coupled with ATP hydrolysis (Fig. 1) and play an essential role in maintaining Ca 2ϩ homeostasis in the cytoplasm and endoplasmic reticulum lumen of cells (1-7). SERCAs have three cytoplasmic domains: phosphorylation (P), nucleotide binding (N), and actuator (A) and 10 transmembrane helices (M1-M10 or 11 in the SERCA2b isoform, M11). In the Ca 2ϩ transport cycle, the ATPase is activated by the binding of two Ca 2ϩ ions from the cytoplasm to the transport sites composed of M4, M5, M6, and M8 (E2 3 E1Ca 2 , step 1). Asp 351 in the P domain is then phosphorylated with MgATP to form the phosphorylated intermediate (EP) (step 2). During dephosphorylation of EP, the Ca 2ϩ ions are released into the lumen. In the detailed mechanism, the dephosphorylation process includes the conformational transition of EP associated with Ca 2ϩ release (step 3) and the subsequent hydrolysis of the acylphosphate bond (step 4).The three human SERCA genes encode SERCA isoforms (8 -10). Mutations in the SERCA2 gene (ATP2A2) and the resulting defects in the SERCA2b housekeeping isoform cause an autosomal dominant genetic skin disease, Darier disease (DD) (11,12). Over 100 mutations have been found with the DD pedigrees (11-24). They include many nonsense mutations, and also substitution and deletion mutations of amino acid residues. The mutations are located throughout the SERCA2b molecule and show no "hot spots" on the primary sequence. To understand how each of the substitution and deletion mutations affects SERCA2b protein, a limited number of mutations had been explored (9 by Ahn et al. (25), 10 by Dode et al. (23), 3 by Sato et al. (26) (a total of 20 because of overlap in Refs. 23 and 25)). To provide a comprehensive insight into the molecular basis of DD, as well as to understand the basis for each case of the DD pedigrees, it is necessary to analyze further the many unexplored substitution and deletion mutations. We therefore carried out in this study a comprehensive analysis of the expression and function of most of the DD causing substitution and deletion mutations reported, i.e. the 51 mutations shown in Fig. 2. Our results showed that most of the mutations (48 of the 51) cause severe defects in protein expression and/or Ca 2ϩ transport function. The loss of the transport function was ascribed to markedly reduced ATP hydrolysis or uncoupling from ATP hydrolysis. The remaining three mutations were exceptional in that they exhibited seemingly normal protein expression and

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Quasi-irreversible inactivation of the sarcoplasmic reticulum Ca2+-ATPase by simultaneous tight binding of magnesium and fluoride to the catalytic site

Cited by 31 publications

References 44 publications

Stable Structural Analog of Ca2+-ATPase ADP-insensitive Phosphoenzyme with Occluded Ca2+ Formed by Elongation of A-domain/M1′-linker and Beryllium Fluoride Binding

Stable Structural Analog of Ca2+-ATPase ADP-insensitive Phosphoenzyme with Occluded Ca2+ Formed by Elongation of A-domain/M1′-linker and Beryllium Fluoride Binding

Concerted Conformational Effects of Ca²⁺and ATP Are Required for Activation of Sequential Reactions in the Ca²⁺ATPase (SERCA) Catalytic Cycle^,

Comprehensive Analysis of Expression and Function of 51 Sarco(endo)plasmic Reticulum Ca2+-ATPase Mutants Associated with Darier Disease

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Quasi-irreversible inactivation of the sarcoplasmic reticulum Ca2+-ATPase by simultaneous tight binding of magnesium and fluoride to the catalytic site

Cited by 31 publications

References 44 publications

Stable Structural Analog of Ca2+-ATPase ADP-insensitive Phosphoenzyme with Occluded Ca2+ Formed by Elongation of A-domain/M1′-linker and Beryllium Fluoride Binding

Stable Structural Analog of Ca2+-ATPase ADP-insensitive Phosphoenzyme with Occluded Ca2+ Formed by Elongation of A-domain/M1′-linker and Beryllium Fluoride Binding

Concerted Conformational Effects of Ca2+and ATP Are Required for Activation of Sequential Reactions in the Ca2+ATPase (SERCA) Catalytic Cycle,

Comprehensive Analysis of Expression and Function of 51 Sarco(endo)plasmic Reticulum Ca2+-ATPase Mutants Associated with Darier Disease

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Concerted Conformational Effects of Ca²⁺and ATP Are Required for Activation of Sequential Reactions in the Ca²⁺ATPase (SERCA) Catalytic Cycle^,