Quantitative analysis of intracellular Nτ-methylhistidine concentration in C2C12 myotubes by liquid chromatography-tandem mass spectrometry.

IF 1.4 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioscience, Biotechnology, and Biochemistry Pub Date : 2025-05-21 DOI:10.1093/bbb/zbaf031

Saki Shimamoto, Miyu Kamimura, Yoshio Nishimura, Jun-Ichi Shiraishi, Shozo Tomonaga, Hanwool Do, Ayumi Katafuchi, Susumu Muroya, Akira Ohtsuka, Daichi Ijiri

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引用次数: 0

Abstract

The release of Nτ-methylhistidine from muscle cells into the culture medium is considered an index of muscle protein breakdown. The aim of this study was to establish a quantitative method for measuring the intercellular concentrations of Nτ-methylhistidine and its isomer Nπ-methylhistidine in C2C12 myotubes via liquid chromatography-tandem mass spectrometry with stable isotope dilution analysis. The acceptable linear ranges of detection were both 0.5-250 pmol/mL; r2 = 1.000 (Nτ-methylhistidine), r2 = 0.999 (Nπ-methylhistidine). Culture media and cell extracts from C2C12 myotubes grown in a 6-well plate were considered acceptable samples for detecting the concentration of Nτ-methylhistidine. However, Nπ-methylhistidine was detected in neither the culture medium nor cell extracts. The proposed method can be used to confirm the effects of amino acid depletion on Nτ-methylhistidine levels in C2C12 myotubes. In C2C12 myotubes, culture in an amino acid-depleted medium for 48 h increased intracellular Nτ-methylhistidine levels while decreasing its extracellular level.

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液相色谱-串联质谱法定量分析C2C12肌管细胞内n τ-甲基组氨酸浓度。

从肌肉细胞释放出新τ-甲基组氨酸到培养基中被认为是肌肉蛋白分解的一个指标。本研究建立了一种液相色谱-串联质谱法测定C2C12肌管中n π-甲基组氨酸及其异构体n π-甲基组氨酸细胞间浓度的定量方法。可接受的线性检测范围为0.5 ~ 250 pmol/mL；r2 = 1.000 （n τ-甲基组氨酸），r2 = 0.999 （n π-甲基组氨酸）。6孔板培养的C2C12肌管培养基和细胞提取物被认为是检测n τ-甲基组氨酸浓度的可接受样品。但在培养基和细胞提取物中均未检测到n π-甲基组氨酸。该方法可用于证实氨基酸缺失对C2C12肌管中n τ-甲基组氨酸水平的影响。在C2C12肌管中，在缺乏氨基酸的培养基中培养48小时，细胞内n τ-甲基组氨酸水平升高，细胞外n τ-甲基组氨酸水平降低。

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

Bioscience, Biotechnology, and Biochemistry 生物-生化与分子生物学

CiteScore

3.50

自引率

0.00%

发文量

183

审稿时长

1 months

期刊介绍： Bioscience, Biotechnology, and Biochemistry publishes high-quality papers providing chemical and biological analyses of vital phenomena exhibited by animals, plants, and microorganisms, the chemical structures and functions of their products, and related matters. The Journal plays a major role in communicating to a global audience outstanding basic and applied research in all fields subsumed by the Japan Society for Bioscience, Biotechnology, and Agrochemistry (JSBBA).