Measurement of Sarcoplasmic Reticulum Ca2+-ATPase Activity and E-Type Mg2+-ATPase Activity in Rat Heart Homogenates

Saborido, Ana; Delgado, Jerónimo; Megías, Alicia

doi:10.1006/abio.1998.3043

Cited by 21 publications

(14 citation statements)

References 26 publications

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“…The SERCA activity measured in Drosophila microsomal membranes was in the same range as those reported elsewhere for vertebrate isoforms (Saborido et al, 1999;Chu et al, 1988). Thapsigargin is a specific inhibitor of SERCAs, and has been used as a pharmacological probe to detect this family of enzymes.…”

Section: Discussionsupporting

confidence: 65%

“…The Mg 2+ -ATPase activity measured according to Saborido et al (Saborido et al, 1999), is designed to evaluate the activity of E-type ATPases, and so it could be that the activity we measured in the Drosophila microsomal fraction is of E-type. Additional experiments are necessary to characterize the sensitivity of this enzymatic activity to specific inhibitors and substrates, to compare SERCA with other E-type Mg 2+ -ATPases (Plesner, 1995).…”

Section: Discussionmentioning

confidence: 99%

“…Measurement of Ca 2+ and Mg 2+ ATPases activities ATPase activities were measured following Saborido et al (Saborido et al, 1999), by using the coupled enzymatic assay of Chu et al (Chu et al, 1988), where the rate of ATP hydrolysis is calculated from the spectrophotometric data of NADH oxidation at 340 nm.…”

Section: Cellular Fractionsmentioning

confidence: 99%

“…3). We employed the procedure reported by Saborido et al (Saborido et al, 1999). First, so-called 'total' ATPase activity was determined: Mainly Mg 2+ and Ca 2+ -ATPase activities.…”

Section: [ 3 H]-ryanodine-binding Assaysmentioning

confidence: 99%

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Biochemical characterization, distribution and phylogenetic analysis ofDrosophila melanogasterryanodine and IP3 receptors, and thapsigargin-sensitive Ca2+ ATPase

Vázquez‐Martínez

Cañedo-Merino

Dı́az-Muñoz

et al. 2003

Journal of Cell Science

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IntroductionIntracellular Ca 2+ dynamics is a key factor in cellular signaling and physiology (for a review, see Bootman et al., 2001). In particular, the system of endomembranes that forms the sarco(endo)plasmic reticulum plays a vital role in Ca 2+ handling in most eukaryots (Carafoli and Klee, 1999). In this compartment two families of intracellular Ca 2+ release channels have been characterized: the ryanodine receptors (RyR) and the inositol 1,4,5-trisphosphate receptors (IP3R) (Nori et al., 1993). Besides these, there is evidence for two more intracellular Ca 2+ releasing channels: the NAAPD and the sphingolipid receptors (Petersen and Cancela, 1999;Cancela, 2001). Ca 2+ re-uptake by the sarco(endo)plasmic reticulum is mediated by the thapsigargin-sensitive sarco(endo)plasmic reticulum Ca 2+ ATPase (SERCA), whose structure and function has been studied extensively in mammalian muscle systems (MacLennan, 1990).The development of the fruit fly Drosophila melanogaster is amenable to multidisciplinary analyses (for a review, see Campos-Ortega and Hartenstein, 1997) and is thus a powerful system in which to examine the role of these proteins in intracellular Ca 2+ homeostasis. In this organism, a single RyR gene with 26 exons, dry, and a single IP3R gene with 12 exons, dip, exist and have been genetically characterized (Takeshima et al., 1994;Sinha and Hasan, 1999). In contrast, RyR and IP3R in vertebrates are coded by at least three different genes that, due to alternative splicing, present a large number of isoforms (Rubtsov and Batrukova, 1997;Marks, 1997). A similar situation occurs with the thapsigargin-sensitive Ca 2+ ATPase. In D. melanogaster only one gene coding for this P-type ATPase has been detected, CaP60A (Magyar et al., 1995); whereas in vertebrates, at least three different isoforms of this Ca 2+ ATPase have been reported (Misquitta et al., 1999).Despite extensive genetic and molecular biology data for these proteins, there is a dearth of basic biochemical information on the Drosophila RyR, IP3R and SERCA proteins. In order to reap the benefit from a genetic and molecular biology tractable model organism with single RyR, IP 3 R and SERCA proteins, we characterize here these important molecules using Drosophila native endomembranes.

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Section: Discussionsupporting

confidence: 65%

Section: Discussionmentioning

confidence: 99%

Section: Cellular Fractionsmentioning

confidence: 99%

“…3). We employed the procedure reported by Saborido et al (Saborido et al, 1999). First, so-called 'total' ATPase activity was determined: Mainly Mg 2+ and Ca 2+ -ATPase activities.…”

Section: [ 3 H]-ryanodine-binding Assaysmentioning

confidence: 99%

See 2 more Smart Citations

Biochemical characterization, distribution and phylogenetic analysis ofDrosophila melanogasterryanodine and IP3 receptors, and thapsigargin-sensitive Ca2+ ATPase

Vázquez‐Martínez

Cañedo-Merino

Dı́az-Muñoz

et al. 2003

Journal of Cell Science

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“…The major fraction (≥80 %) was unstable decaying exponentially with a T 1/2 of 2-3 min and could be attributed to NTPDase2. This assumption is based on ATPdependent inactivation reported first for some rat Mg-ATPases or ecto-ATPases [27][28][29][30] and more recently, since NTPDases have been identified and cloned, for human NTPDase2 [31]. The stable (final) ATPase activity was similar to ADPase activity and could be due to NTPDase1, NTPDase3, or NTPDase8.…”

Section: Resultsmentioning

confidence: 99%

Rat submandibular glands secrete nanovesicles with NTPDase and 5′-nucleotidase activities

González

Egido

Balcarcel

et al. 2014

Purinergic Signalling

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Extracellular nucleotides modulate a wide number of biological processes such as neurotransmission, platelet aggregation, muscle contraction, and epithelial secretion acting by the purinergic pathway. Nucleotidases as NTPDases and ecto-5′-nucleotidase are membraneanchored proteins that regulate extracellular nucleotide concentrations. In a previous work, we have partially characterized an NTPDase-like activity expressed by rat submandibular gland microsomes, giving rise to the hypothesis that membrane NTPDases could be released into salivary ducts to regulate luminal nucleotide concentrations as was previously proposed for ovarian, prostatic, and pancreatic secretions. Present results show that rat submandibular glands incubated in vitro release membrane-associated NTPDase and ecto-5′-nucleotidase activities. Electron microscopy images show that released membranes presenting nucleotidase activity correspond to exosome-like vesicles which are also present at microsomal fraction. Both exosome release and nucleotidase activities are raised by adrenergic stimulation. Nucleotidase activities present the same kinetic characteristics than microsomal nucleotidase activity, corresponding mainly to the action of NTPDase2 and NTPDase3 isoforms as well as 5′-nucleotidase. This is consistent with Western blot analysis revealing the presence of these enzymes in the microsomal fraction.

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Two different types of kinetics, where the initial rate increases faster or slower than the reactant concentration, can coexist on bell-shaped kinetic dependencies

Zhuk

Karakhim

2023

J Math Chem

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Measurement of Sarcoplasmic Reticulum Ca2+-ATPase Activity and E-Type Mg2+-ATPase Activity in Rat Heart Homogenates

Cited by 21 publications

References 26 publications

Biochemical characterization, distribution and phylogenetic analysis ofDrosophila melanogasterryanodine and IP3 receptors, and thapsigargin-sensitive Ca2+ ATPase

Biochemical characterization, distribution and phylogenetic analysis ofDrosophila melanogasterryanodine and IP3 receptors, and thapsigargin-sensitive Ca2+ ATPase

Rat submandibular glands secrete nanovesicles with NTPDase and 5′-nucleotidase activities

Two different types of kinetics, where the initial rate increases faster or slower than the reactant concentration, can coexist on bell-shaped kinetic dependencies

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