2010
DOI: 10.1042/bj20091956
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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 loca… Show more

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Cited by 38 publications
(38 citation statements)
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“…From these data, it was proposed that, in the sea urchin egg, the primary targets of NAADP are acidic stores, probably lysosome-related organelles. Moreover, a recent study has ruled out polyphosphate-containing acidocalcisomes [27]. Intriguingly, experiments in sea urchin egg homogenates employing luminal pH indicators such as Acridine Orange or LysoSensor TM also have demonstrated that NAADP, but not InsP 3 or cADPR, also causes the alkalinization of acidic stores, which may be an additional important aspect of NAADP-mediated signalling mechanisms [28].…”
Section: Naadp-sensitive Ca 2+ Storesmentioning
confidence: 97%
“…From these data, it was proposed that, in the sea urchin egg, the primary targets of NAADP are acidic stores, probably lysosome-related organelles. Moreover, a recent study has ruled out polyphosphate-containing acidocalcisomes [27]. Intriguingly, experiments in sea urchin egg homogenates employing luminal pH indicators such as Acridine Orange or LysoSensor TM also have demonstrated that NAADP, but not InsP 3 or cADPR, also causes the alkalinization of acidic stores, which may be an additional important aspect of NAADP-mediated signalling mechanisms [28].…”
Section: Naadp-sensitive Ca 2+ Storesmentioning
confidence: 97%
“…This technique was used before to localize polyP in Saccharomyces cerevisiae vacuoles (28) and in acidocalcisomes of sea urchin eggs (49) and Rhodnius prolixus (50). The negative staining by DAPI used for the urea-PAGE analysis allows the discrimination among DNA, RNA, and glycosaminoglycans which do not photobleach in the time frame necessary for complete photobleaching of even nanomolar amounts of polyP, and heparin, which photobleach very slowly on exposure to UV light (32).…”
Section: Discussionmentioning
confidence: 99%
“…25 A potential drawback to using a relative nonspecific enzyme, such as alkaline phosphatase, is that it can also enzymatically degrade other phosphate-containing compounds, such as ADP, an important platelet agonist. And finally, the isolated polyP binding domain of E coli exopolyphosphatase has been used successfully as a probe to localize polyP in yeast cells 107 and in acidocalcisomes in eggs of sea urchins 108 and insects. 109 For a variety of experiments, it would be desirable to be able to covalently attach biotin, epitope tags, dyes, fluorophores, etc, to polyP, and also to covalently immobilize polyP onto solid supports, such as magnetic beads and multiwell plates.…”
Section: Methods For Preparing and Analyzing Polypmentioning
confidence: 99%