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Biochem J
2010 Aug 01;4293:485-95. doi: 10.1042/BJ20091956.
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Calcium- and polyphosphate-containing acidic granules of sea urchin eggs are similar to acidocalcisomes, but are not the targets for NAADP.
Ramos IB
,
Miranda K
,
Pace DA
,
Verbist KC
,
Lin FY
,
Zhang Y
,
Oldfield E
,
Machado EA
,
De Souza W
,
Docampo R
.
Abstract
Acidocalcisomes are acidic calcium-storage compartments described from bacteria to humans and characterized by their high content in poly P (polyphosphate), a linear polymer of many tens to hundreds of Pi residues linked by high-energy phosphoanhydride bonds. In the present paper we report that millimolar levels of short-chain poly P (in terms of Pi residues) and inorganic PPi are present in sea urchin extracts as detected using 31P-NMR, enzymatic determinations and agarose gel electrophoresis. Poly P was localized to granules randomly distributed in the sea urchin eggs, as shown by labelling with the poly-P-binding domain of Escherichia coli exopolyphosphatase. These granules were enriched using iodixanol centrifugation and shown to be acidic and to contain poly P, as determined by Acridine Orange and DAPI (4'',6''-diamidino-2-phenylindole) staining respectively. These granules also contained large amounts of calcium, sodium, magnesium, potassium and zinc, as detected by X-ray microanalysis, and bafilomycin A1-sensitive ATPase, pyrophosphatase and exopolyphosphatase activities, as well as Ca2+/H+ and Na+/H+ exchange activities, being therefore similar to acidocalcisomes described in other organisms. Calcium release from these granules induced by nigericin was associated with poly P hydrolysis. Although NAADP (nicotinic acid-adenine dinucleotide phosphate) released calcium from the granule fraction, this activity was not significantly enriched as compared with the NAADP-stimulated calcium release from homogenates and was not accompanied by poly P hydrolysis. GPN (glycyl-L-phenylalanine-naphthylamide) released calcium when added to sea urchin homogenates, but was unable to release calcium from acidocalcisome-enriched fractions, suggesting that these acidic stores are not the targets for NAADP.
Figure 1. Transmission electron microscopy of A. punctulata eggsThe preparation was fixed and processed for standard electron microscopy. Cortical granules (cg, thin arrow), yolk platelets (YP) and âclear granulesâ containing residual electron-dense material (thick arrow), smaller clear vesicles (white arrowheads) and mitochondria (black arrowheads) are indicated. Scale bars in (A) are 3 μm, (B) 1 μm and (CâE) 0.5 μm.
Figure 2. Electron microscopy and X-ray microanalysis of whole intact eggs and homogenates(A and B) Electron energy-filtered transmission electron microscopy of an unfixed and unstained egg (A) or an egg homogenate (B) from A. punctulata. Dense granules are identified by black arrows. (C) Typical X-ray microanalysis spectrum of dense granules found in egg homogenates [black arrow in (B)]. (D) Typical X-ray microanalysis spectrum of a less-electron-dense vesicle (probably a yolk platelet) [white arrow in (B)] found in egg homogenates. Scale bars in (A) and (B) are 3 μm.
Figure 3. Detection of PPi and poly P in sea urchin eggs(A) PPi, SC poly P and LC poly P were assayed as described in the Materials and methods section, and levels are expressed as mM poly P (in terms of Pi residues). Data are from four experiments, and show means±S.E.M. Different letters indicate significant differences (one-way ANOVA, P<0.05). (B) Agarose gel electrophoresis of poly P stained with Toluidine Blue. Lane 1, poly P extract; lane 2, poly P extract after rPPX1 treatment. P75, P45 and P24 are poly P standards of 75+, 45 and 25 phosphate units. (C) 242.8-MHz (1H decoupled) 31P-NMR spectra of a perchloric acid extract of sea urchins eggs. See assignments in Table 1. (D) Differential interferential microscopy of the sea urchin egg shown in (E). (E) Localization of poly P using the recombinant PPBD of E. coli PPX linked with an Xpress epitope tag. Images were deconvolved for 15 cycles using Softwarx deconvolution software. Other conditions are described in the Materials and methods. Scale bars in (D) and (E) are 15 μm.
AFZELIUS,
The ultrastructure of the cortical granules and their products in the sea urchin egg as studied with the electron microscope.
1956, Pubmed,
Echinobase
AFZELIUS,
The ultrastructure of the cortical granules and their products in the sea urchin egg as studied with the electron microscope.
1956,
Pubmed
,
Echinobase
Ault-Riché,
Novel assay reveals multiple pathways regulating stress-induced accumulations of inorganic polyphosphate in Escherichia coli.
1998,
Pubmed
Brailoiu,
Essential requirement for two-pore channel 1 in NAADP-mediated calcium signaling.
2009,
Pubmed
Brailoiu,
Nicotinic acid adenine dinucleotide phosphate potentiates neurite outgrowth.
2005,
Pubmed
Brailoiu,
An ancestral deuterostome family of two-pore channels mediates nicotinic acid adenine dinucleotide phosphate-dependent calcium release from acidic organelles.
2010,
Pubmed
,
Echinobase
Calcraft,
NAADP mobilizes calcium from acidic organelles through two-pore channels.
2009,
Pubmed
Cancela,
Coordination of agonist-induced Ca2+-signalling patterns by NAADP in pancreatic acinar cells.
1999,
Pubmed
,
Echinobase
Churchill,
NAADP mobilizes Ca(2+) from reserve granules, lysosome-related organelles, in sea urchin eggs.
2002,
Pubmed
,
Echinobase
Churchill,
NAADP induces Ca2+ oscillations via a two-pool mechanism by priming IP3- and cADPR-sensitive Ca2+ stores.
2001,
Pubmed
,
Echinobase
Clapper,
Pyridine nucleotide metabolites stimulate calcium release from sea urchin egg microsomes desensitized to inositol trisphosphate.
1987,
Pubmed
,
Echinobase
Docampo,
Acidocalcisomes - conserved from bacteria to man.
2005,
Pubmed
Epel,
Molecular mechanisms for prevention of polyspermy.
1975,
Pubmed
,
Echinobase
Jadot,
Intralysosomal hydrolysis of glycyl-L-phenylalanine 2-naphthylamide.
1984,
Pubmed
Johnson,
Intracellular pH and activation of sea urchin eggs after fertilisation.
1976,
Pubmed
,
Echinobase
Lanzetta,
An improved assay for nanomole amounts of inorganic phosphate.
1979,
Pubmed
Lee,
Functional visualization of the separate but interacting calcium stores sensitive to NAADP and cyclic ADP-ribose.
2000,
Pubmed
,
Echinobase
Lee,
A derivative of NADP mobilizes calcium stores insensitive to inositol trisphosphate and cyclic ADP-ribose.
1995,
Pubmed
,
Echinobase
Lee,
Changes in intracellular acidic compartments in sea urchin eggs after activation.
1983,
Pubmed
,
Echinobase
Lemercier,
A pyrophosphatase regulating polyphosphate metabolism in acidocalcisomes is essential for Trypanosoma brucei virulence in mice.
2004,
Pubmed
Lu,
Ca2+ content and expression of an acidocalcisomal calcium pump are elevated in intracellular forms of Trypanosoma cruzi.
1998,
Pubmed
Maeshima,
TONOPLAST TRANSPORTERS: Organization and Function.
2001,
Pubmed
Mallya,
Proteolysis of the major yolk glycoproteins is regulated by acidification of the yolk platelets in sea urchin embryos.
1992,
Pubmed
,
Echinobase
Mándi,
Ca2+ release triggered by NAADP in hepatocyte microsomes.
2006,
Pubmed
,
Echinobase
Marchesini,
Acidocalcisomes are functionally linked to the contractile vacuole of Dictyostelium discoideum.
2002,
Pubmed
Mitchell,
Ryanodine receptor type I and nicotinic acid adenine dinucleotide phosphate receptors mediate Ca2+ release from insulin-containing vesicles in living pancreatic beta-cells (MIN6).
2003,
Pubmed
Morgan,
Fertilization and nicotinic acid adenine dinucleotide phosphate induce pH changes in acidic Ca(2+) stores in sea urchin eggs.
2007,
Pubmed
,
Echinobase
Morgan,
NAADP induces pH changes in the lumen of acidic Ca2+ stores.
2007,
Pubmed
,
Echinobase
Motta,
Proton-pyrophosphatase and polyphosphate in acidocalcisome-like vesicles from oocytes and eggs of Periplaneta americana.
2009,
Pubmed
Parrington,
Flipping the switch: how a sperm activates the egg at fertilization.
2007,
Pubmed
,
Echinobase
Patel,
Acidic calcium stores open for business: expanding the potential for intracellular Ca2+ signaling.
2010,
Pubmed
Ramos,
Calcium- and polyphosphate-containing acidocalcisomes in chicken egg yolk.
2010,
Pubmed
Ratto,
Purification and characterization of arginine kinase from sea-urchin eggs.
1988,
Pubmed
,
Echinobase
Rodrigues,
Characterization of a vacuolar pyrophosphatase in Trypanosoma brucei and its localization to acidocalcisomes.
1999,
Pubmed
Rodrigues,
An acidocalcisomal exopolyphosphatase from Leishmania major with high affinity for short chain polyphosphate.
2002,
Pubmed
Rodrigues,
Presence of a vacuolar H+-pyrophosphatase in promastigotes of Leishmania donovani and its localization to a different compartment from the vacuolar H+-ATPase.
1999,
Pubmed
Ruiz,
The polyphosphate bodies of Chlamydomonas reinhardtii possess a proton-pumping pyrophosphatase and are similar to acidocalcisomes.
2001,
Pubmed
Ruiz,
Human platelet dense granules contain polyphosphate and are similar to acidocalcisomes of bacteria and unicellular eukaryotes.
2004,
Pubmed
Ruiz,
Rapid changes in polyphosphate content within acidocalcisomes in response to cell growth, differentiation, and environmental stress in Trypanosoma cruzi.
2001,
Pubmed
Saito,
Direct labeling of polyphosphate at the ultrastructural level in Saccharomyces cerevisiae by using the affinity of the polyphosphate binding domain of Escherichia coli exopolyphosphatase.
2005,
Pubmed
Sardet,
The ultrastructure of the sea urchin egg cortex isolated before and after fertilization.
1984,
Pubmed
,
Echinobase
Seufferheld,
The H(+)-pyrophosphatase of Rhodospirillum rubrum is predominantly located in polyphosphate-rich acidocalcisomes.
2004,
Pubmed
Seufferheld,
Identification of organelles in bacteria similar to acidocalcisomes of unicellular eukaryotes.
2003,
Pubmed
Shen,
Direct measurement of intracellular pH during metabolic derepression of the sea urchin egg.
1978,
Pubmed
,
Echinobase
Steen,
NAADP mobilizes calcium from the endoplasmic reticular Ca(2+) store in T-lymphocytes.
2007,
Pubmed
Terasaki,
Labeling of cell membranes and compartments for live cell fluorescence microscopy.
2004,
Pubmed
Vercesi,
Sodium-proton exchange stimulates Ca2+ release from acidocalcisomes of Trypanosoma brucei.
1996,
Pubmed
Vercesi,
Presence of a Na(+)/H(+) exchanger in acidocalcisomes of Leishmania donovani and their alkalization by anti-leishmanial drugs.
2000,
Pubmed
Yamasaki,
Role of NAADP and cADPR in the induction and maintenance of agonist-evoked Ca2+ spiking in mouse pancreatic acinar cells.
2005,
Pubmed
,
Echinobase
Zhang,
Production of NAADP and its role in Ca2+ mobilization associated with lysosomes in coronary arterial myocytes.
2006,
Pubmed
Zhang,
Reconstitution and characterization of a nicotinic acid adenine dinucleotide phosphate (NAADP)-sensitive Ca2+ release channel from liver lysosomes of rats.
2007,
Pubmed
Zong,
The two-pore channel TPCN2 mediates NAADP-dependent Ca(2+)-release from lysosomal stores.
2009,
Pubmed