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非变性聚丙烯酰胺凝胶电泳分析植物 L-半胱氨酸脱硫酶(LCD)同工酶。

Analysis of Plant L-Cysteine Desulfhydrase (LCD) Isozymes by Non-denaturing Polyacrylamide Gel Electrophoresis.

机构信息

Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Stress, Development and Signaling in Plants, Estación Experimental del Zaidín, Spanish National Research Council (CSIC), Granada, Spain.

出版信息

Methods Mol Biol. 2023;2642:233-240. doi: 10.1007/978-1-0716-3044-0_13.

Abstract

Hydrogen sulfide (HS) is a signaling molecule that achieves different regulatory functions in animal and plant cells. The cytosolic enzyme L-cysteine desulfhydrase (LCD; EC 4.4.1.28) catalyzes the conversion of cysteine (L-Cys) to pyruvate and ammonium with the concomitant generation of HS, this enzyme being considered one of the main sources of HS in higher plants. Using non-denaturing polyacrylamide gel electrophoresis (PAGE) in combination with a specific assay for LCD activity, the present protocol allows identifying diverse LCD isozymes present in different organs (roots, shoots, leaves, and fruits) and plant species including pea, garlic, Arabidopsis, and pepper.

摘要

硫化氢 (HS) 是一种信号分子,在动物和植物细胞中实现不同的调节功能。胞质酶 L-半胱氨酸脱硫酶 (LCD;EC 4.4.1.28) 催化半胱氨酸 (L-Cys) 转化为丙酮酸和铵,同时生成 HS,该酶被认为是高等植物中 HS 的主要来源之一。本协议使用非变性聚丙烯酰胺凝胶电泳 (PAGE) 结合用于 LCD 活性的特定测定法,允许鉴定存在于不同器官(根、茎、叶和果实)和包括豌豆、大蒜、拟南芥和辣椒在内的植物物种中的各种 LCD 同工酶。

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