An Improved Two-Dimensional HPLC Method for Endogenous 2'-Deoxy-ADPR and 2'-Deoxy-NAD.

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4414-0_6

Feng Gu, Ralf Fliegert, Andreas Bauche, Andreas H Guse

引用次数: 0

Abstract

Transient receptor potential melastatin 2 (TRPM2) is a multifunctional non-selective Ca²⁺-permeable cation channel expressed in numerous immune cells. 2'-Deoxy-adenosine diphosphoribose (2d-ADPR) has been identified as a superagonist of TRPM2 channels that induces higher whole-cell currents and requires a lower intracellular Ca²⁺ concentration for activation. 2d-ADPR can be produced from 2'-deoxy-nicotinamide adenine dinucleotide (2d-NAD) by the enzyme CD38 in vitro under physiological conditions.Here, we describe a two-dimensional HPLC method, suitable for the quantification of endogenous 2d-ADPR and 2d-NAD. The results demonstrated that 25 million Jurkat T cells per sample were sufficient to quantify endogenous 2d-ADPR and 2d-NAD.

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内源性2’-脱氧- adpr和2’-脱氧- nad的改进二维高效液相色谱法

瞬时受体电位美拉他汀2 （TRPM2）是一种多功能的非选择性Ca2+渗透性阳离子通道，在许多免疫细胞中表达。2'-脱氧腺苷二磷酸核糖（2d-ADPR）已被确定为TRPM2通道的超级激动剂，可诱导更高的全细胞电流，并且需要较低的细胞内Ca2+浓度才能激活。2'-脱氧烟酰胺腺嘌呤二核苷酸（2d-NAD）在体外生理条件下可由CD38酶合成2d-ADPR。本文建立了一种适用于内源性2d-ADPR和2d-NAD定量的二维高效液相色谱方法。结果表明，每个样本2500万个Jurkat T细胞足以量化内源性2d-ADPR和2d-NAD。

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 in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.