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克氏锥虫酸性钙小体中多磷酸盐含量随细胞生长、分化及环境应激的快速变化

Rapid changes in polyphosphate content within acidocalcisomes in response to cell growth, differentiation, and environmental stress in Trypanosoma cruzi.

作者信息

Ruiz F A, Rodrigues C O, Docampo R

机构信息

Laboratory of Molecular Parasitology, Department of Pathobiology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802, USA.

出版信息

J Biol Chem. 2001 Jul 13;276(28):26114-21. doi: 10.1074/jbc.M102402200. Epub 2001 May 22.

Abstract

Inorganic polyphosphate (polyP) has been identified and measured in different stages of Trypanosoma cruzi. Millimolar levels (in terms of P(i) residues) in chains of less than 50 residues long, and micromolar levels in chains of about 700--800 residues long, were found in different stages of T. cruzi. Analysis of purified T. cruzi acidocalcisomes indicated that polyPs were preferentially located in these organelles. This was confirmed by visualization of polyPs in the acidocalcisomes using 4',6-diamidino-2-phenylindole. A rapid increase (within 2--4 h) in the levels of short and long chain polyPs was detected during trypomastigote to amastigote differentiation and during the lag phase of growth of epimastigotes (within 12--24 h). Levels rapidly decreased after the epimastigotes resumed growth. Short and long chain polyP levels rapidly decreased upon exposure of epimastigotes to hypo-osmotic or alkaline stresses, whereas levels increased after hyperosmotic stress. Ca(2+) release from acidocalcisomes by a combination of ionophores (ionomycin and nigericin) was associated with the hydrolysis of short and long chain polyPs. In agreement with these results, acidocalcisomes were shown to contain polyphosphate kinase and exopolyphosphatase activities. Together, these results suggest a critical role for these organelles in the adaptation of the parasite to environmental changes.

摘要

在克氏锥虫的不同阶段已鉴定并测量了无机多聚磷酸盐(polyP)。在克氏锥虫的不同阶段,发现长度小于50个残基的链中多聚磷酸盐水平为毫摩尔级(以无机磷酸(P(i))残基计),而长度约为700 - 800个残基的链中为微摩尔级。对纯化的克氏锥虫酸性钙小体的分析表明,多聚磷酸盐优先位于这些细胞器中。使用4',6-二脒基-2-苯基吲哚对酸性钙小体中的多聚磷酸盐进行可视化证实了这一点。在锥鞭毛体向无鞭毛体分化期间以及上鞭毛体生长的延迟期(12 - 24小时内),检测到短链和长链多聚磷酸盐水平迅速增加(2 - 4小时内)。上鞭毛体恢复生长后,其水平迅速下降。上鞭毛体暴露于低渗或碱性应激时,短链和长链多聚磷酸盐水平迅速下降,而在高渗应激后水平升高。离子载体(离子霉素和尼日利亚菌素)组合导致酸性钙小体释放Ca(2+)与短链和长链多聚磷酸盐的水解有关。与这些结果一致,酸性钙小体显示含有多聚磷酸激酶和外多聚磷酸酶活性。总之,这些结果表明这些细胞器在寄生虫适应环境变化中起关键作用。

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