دورية أكاديمية
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E2P-like states of plasma membrane Ca 2+ ‑ATPase characterization of vanadate and fluoride-stabilized phosphoenzyme analogues.

التفاصيل البيبلوغرافية
العنوان: E2P-like states of plasma membrane Ca 2+ ‑ATPase characterization of vanadate and fluoride-stabilized phosphoenzyme analogues.
المؤلفون: Saffioti NA; Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Instituto de Química y Fisicoquímica Biológicas (IQUIFIB), Facultad de Farmacia y Bioquímica, Junín 956, Ciudad Autónoma de Buenos Aires C1113AAD, Argentina., de Sautu M; Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Instituto de Química y Fisicoquímica Biológicas (IQUIFIB), Facultad de Farmacia y Bioquímica, Junín 956, Ciudad Autónoma de Buenos Aires C1113AAD, Argentina., Ferreira-Gomes MS; Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Instituto de Química y Fisicoquímica Biológicas (IQUIFIB), Facultad de Farmacia y Bioquímica, Junín 956, Ciudad Autónoma de Buenos Aires C1113AAD, Argentina., Rossi RC; Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Instituto de Química y Fisicoquímica Biológicas (IQUIFIB), Facultad de Farmacia y Bioquímica, Junín 956, Ciudad Autónoma de Buenos Aires C1113AAD, Argentina., Berlin J; Department of Pharmacology and Physiology, New Jersey Medical School, Rutgers University, Newark, NJ, USA., Rossi JPFC; Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Instituto de Química y Fisicoquímica Biológicas (IQUIFIB), Facultad de Farmacia y Bioquímica, Junín 956, Ciudad Autónoma de Buenos Aires C1113AAD, Argentina., Mangialavori IC; Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Instituto de Química y Fisicoquímica Biológicas (IQUIFIB), Facultad de Farmacia y Bioquímica, Junín 956, Ciudad Autónoma de Buenos Aires C1113AAD, Argentina. Electronic address: irenem@qb.ffyb.uba.ar.
المصدر: Biochimica et biophysica acta. Biomembranes [Biochim Biophys Acta Biomembr] 2019 Feb 01; Vol. 1861 (2), pp. 366-379. Date of Electronic Publication: 2018 Nov 09.
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Elsevier Country of Publication: Netherlands NLM ID: 101731713 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-2642 (Electronic) Linking ISSN: 00052736 NLM ISO Abbreviation: Biochim Biophys Acta Biomembr Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Amsterdam : Elsevier
مواضيع طبية MeSH: Calcium-Transporting ATPases/*chemistry , Calcium-Transporting ATPases/*metabolism , Cell Membrane/*enzymology , Fluorides/*pharmacology , Vanadates/*pharmacology, Adenosine Triphosphate/metabolism ; Calcium-Transporting ATPases/antagonists & inhibitors ; Calmodulin/metabolism ; Enzyme Stability/drug effects ; Eosine Yellowish-(YS)/metabolism ; Fluorescence ; Humans ; Hydrogen-Ion Concentration ; Kinetics ; Magnesium/pharmacology ; Phosphoprotein Phosphatases/metabolism ; Phosphorylation/drug effects ; Protein Conformation ; Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism ; Time Factors ; Water
مستخلص: The plasma membrane Ca 2+ ‑ATPase (PMCA) belongs to the family of P-type ATPases, which share the formation of an acid-stable phosphorylated intermediate as part of their reaction cycle. The crystal structure of PMCA is currently lacking. Its abundance is approximately 0.1% of the total protein in the membrane, hampering efforts to produce suitable crystals for X-ray structure analysis. In this work we characterized the effect of beryllium fluoride (BeF x ), aluminium fluoride (AlF x ) and magnesium fluoride (MgF x ) on PMCA. These compounds are known inhibitors of P-type ATPases that stabilize E2P ground, E2·P phosphoryl transition and E2·P i product states. Our results show that the phosphate analogues BeF x , AlF x and MgF x inhibit PMCA Ca 2+ ‑ATPase activity, phosphatase activity and phosphorylation with high apparent affinity. Ca 2+ ‑ATPase inhibition by AlF x and BeF x depended on Mg 2+ concentration indicating that this ion stabilizes the complex between these inhibitors and the enzyme. Low pH increases AlF x and BeF x but not MgF x apparent affinity. Eosin fluorescent probe binds with high affinity to the nucleotide binding site of PMCA. The fluorescence of eosin decreases when fluoride complexes bind to PMCA indicating that the environment of the nucleotide binding site is less hydrophobic in E2P-like states. Finally, measuring the time course of E → E2P-like conformational change, we proposed a kinetic model for the binding of fluoride complexes and vanadate to PMCA. In summary, our results show that these fluoride complexes reveal different states of phosphorylated intermediates belonging to the mechanism of hydrolysis of ATP by the PMCA.
(Copyright © 2018. Published by Elsevier B.V.)
فهرسة مساهمة: Keywords: Metal fluorides; PMCA; Phosphorylated state; P‑ATPases; Reaction cycle
المشرفين على المادة: 0 (Calmodulin)
059QF0KO0R (Water)
3WHH0066W5 (Vanadates)
8L70Q75FXE (Adenosine Triphosphate)
EC 3.1.3.16 (Phosphoprotein Phosphatases)
EC 3.6.3.8 (Sarcoplasmic Reticulum Calcium-Transporting ATPases)
EC 7.2.2.10 (Calcium-Transporting ATPases)
I38ZP9992A (Magnesium)
Q80VPU408O (Fluorides)
TDQ283MPCW (Eosine Yellowish-(YS))
تواريخ الأحداث: Date Created: 20181113 Date Completed: 20190919 Latest Revision: 20191210
رمز التحديث: 20231215
DOI: 10.1016/j.bbamem.2018.11.001
PMID: 30419189
قاعدة البيانات: MEDLINE
الوصف
تدمد:1879-2642
DOI:10.1016/j.bbamem.2018.11.001