Optimized biochemical method for human Polyphosphate quantification

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods Pub Date : 2025-02-01 DOI:10.1016/j.ymeth.2025.01.001

Blanca Lázaro , Ana Sarrias , Francisco J. Tadeo , Joan Marc Martínez-Láinez , Ainhoa Fernández , Eva Quandt , Blanca Depares , Tobias Dürr-Mayer , Henning Jessen , Javier Jiménez , Josep Clotet , Samuel Bru

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引用次数: 0

Abstract

Polyphosphate (polyP), a biopolymer composed of phosphates, impacts a wide range of biological functions and pathological conditions in all organisms. However, polyP’s intricate physiology and structure in human cells have remained elusive, largely due to the lack of a reliable quantification method including its extraction. In this study, we assess critical points in the whole process: extraction, purification, and quantification polyP from human cell lines. We developed a highly efficient method that extracts between 3 and 100 times more polyP than previously achieved. Supported by Nuclear Magnetic Resonance (NMR), our approach confirms that mammalian polyP is primarily a linear unbranched polymer. We applied the optimized method to commonly used human cell lines, uncovering important variations of intracellular polyP that correlate with the expression levels of specific polyP converting enzymes. This study underscores the importance of employing several techniques for polyP characterization in parallel and provides a valuable and standardized tool for further exploration in this field.

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优化人多磷酸盐定量的生化方法。

聚磷酸盐（polyP）是一种由磷酸盐组成的生物聚合物，在所有生物体中影响着广泛的生物功能和病理状况。然而，由于缺乏可靠的定量方法，包括其提取，息肉蛋白在人类细胞中复杂的生理和结构仍然是难以捉摸的。在这项研究中，我们评估了整个过程的关键点：从人细胞系中提取、纯化和定量息肉。我们开发了一种高效的方法，提取的息肉比以前多3到100倍。在核磁共振（NMR）的支持下，我们的方法证实了哺乳动物息肉主要是一种线性无支化聚合物。我们将优化的方法应用于常用的人类细胞系，发现细胞内息肉蛋白的重要变化与特定息肉蛋白转化酶的表达水平相关。本研究强调了采用多种技术并行表征息肉的重要性，并为该领域的进一步探索提供了有价值的标准化工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods 生物-生化研究方法

CiteScore

9.80

自引率

2.10%

发文量

222

审稿时长

11.3 weeks

期刊介绍： Methods focuses on rapidly developing techniques in the experimental biological and medical sciences. Each topical issue, organized by a guest editor who is an expert in the area covered, consists solely of invited quality articles by specialist authors, many of them reviews. Issues are devoted to specific technical approaches with emphasis on clear detailed descriptions of protocols that allow them to be reproduced easily. The background information provided enables researchers to understand the principles underlying the methods; other helpful sections include comparisons of alternative methods giving the advantages and disadvantages of particular methods, guidance on avoiding potential pitfalls, and suggestions for troubleshooting.