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采用易环境超声喷雾电离质谱法研究氮饥饿对集胞藻 PCC 6803 膜脂的影响。

The effect of nitrogen starvation on membrane lipids of Synechocystis sp. PCC 6803 investigated by using easy ambient sonic-spray ionization mass spectrometry.

机构信息

Department of Chemistry, University of Crete, Voutes Campus, Heraklion 70013, Greece.

Department of Chemistry, University of Crete, Voutes Campus, Heraklion 70013, Greece.

出版信息

Biochim Biophys Acta Biomembr. 2019 Oct 1;1861(10):183027. doi: 10.1016/j.bbamem.2019.07.015. Epub 2019 Jul 24.

Abstract

BACKGROUND

Glycerolipids are important components of membranes in cyanobacteria that possess vital roles in biological processes. The effect of nitrogen deprivation on membrane lipids of Synechocystis sp. PCC 6803 lipids has not been previously examined.

METHODS

Easy ambient sonic-spray ionization mass spectrometry (EASI-MS) was used for the analysis of Synechocystis 6803 cells with minimal sample preparation, providing rapid qualitative and relative quantitative information on the lipid content of their membranes.

RESULTS

Changes in the degree of unsaturation of membrane lipids were observed for cells grown under normal conditions during different growth phases. This physiological remodeling was disrupted when nitrogen was withdrawn from the cultivation medium. However, this disruption was reversed when the cells were resuspended in normal N-containing medium. Mass spectrometric data were supported by examination of cells by electron microscopy.

CONCLUSIONS

EASI-MS was applied for the first time in the analysis of Synechocystis 6803 cells grown under nitrogen deprived conditions and was found to be a powerful technique operating in a high-throughput manner for the rapid lipid profiling of cells at different growth stages and under different growth conditions.

GENERAL SIGNIFICANCE

The effect of nitrogen deprivation on membrane lipids of Synechocystis 6803 cells was revealed using an ambient ionization technique which enabled high-throughput cell analysis with minimal sample preparation. The results obtained have the potential to be used in future studies to decipher the involvement of enzymes in the observed lipid profile changes.

摘要

背景

甘油酯是蓝藻细胞膜的重要组成部分,在生物过程中起着至关重要的作用。氮饥饿对集胞藻 PCC 6803 脂类膜脂的影响尚未被检测到。

方法

简单环境超声喷雾电离质谱(EASI-MS)用于分析集胞藻 6803 细胞,只需很少的样品制备,即可快速提供有关其膜脂含量的定性和相对定量信息。

结果

在不同的生长阶段,正常条件下生长的细胞的膜脂不饱和程度发生了变化。当从培养介质中去除氮时,这种生理重塑被破坏。然而,当细胞重新悬浮在正常含氮的培养基中时,这种破坏被逆转了。质谱数据得到了电子显微镜检查细胞的支持。

结论

EASI-MS 首次应用于分析在氮饥饿条件下生长的集胞藻 6803 细胞,被发现是一种强大的技术,以高通量的方式对不同生长阶段和不同生长条件下的细胞进行快速脂质分析。

一般意义

用环境离子化技术揭示了氮饥饿对集胞藻 6803 细胞膜脂的影响,该技术能够在最小样品制备的情况下进行高通量细胞分析。所获得的结果有可能在未来的研究中用于破译观察到的脂质图谱变化中酶的参与。

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