Purification of Recombinant Galectins from Different Species Using Distinct Affinity Chromatography Methods.
Purification of Recombinant Galectins from Different Species Using Distinct Affinity Chromatography Methods.
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使用不同的亲和色谱方法纯化不同物种的重组半乳糖素。
DOI：
10.1007/978-1-0716-2055-7_3
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Stowell,SeanR
中科院分区：
文献类型：
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作者：
Paul,Anu;Wu,Shang-Chuen;Patel,KashyapR;Ho,AlexD;Allen,JerryWilliamLynn;Verkerke,Hans;Arthur,ConnieM;Stowell,SeanR
半乳糖凝集素是具有识别含β-半乳糖的聚糖结构的能力的凝集素，并且广泛分布于各种分类群中。然而，确切的生理和生化功能介导的半乳糖凝集素，使其广泛存在于不同的物种还没有被描绘在一个精确的方式。纯化活性形式的重组半乳糖凝集素是阐明其生物学功能的基本要求。在本章中，我们描述了使用三种不同的亲和纯化方法重组表达和纯化半乳糖凝集素的方法，即，乳糖基-琼脂糖色谱法用于真菌半乳糖凝集素灰拟鬼伞半乳糖凝集素2（CGL 2），镍-色谱法用于组氨酸标记的人半乳糖凝集素-7，谷胱甘肽-琼脂糖色谱法用于谷胱甘肽S-转移酶标记（GST-标记）的人半乳糖凝集素-7。提供了用于获得保留碳水化合物结合活性并且适合于进行生物化学实验的上述重组半乳糖凝集素的逐步说明。
Galectins are lectins having the capacity to recognize β-galactose-containing glycan structures and are widely distributed among various taxa. However, the exact physiological and biochemical functions mediated by galectins that necessitate their wide occurrence among diverse species have not yet been delineated in a precise manner. Purification of recombinant galectins in active form is a fundamental requirement to elucidate their biological function. In this chapter, we are describing methods to recombinantly express and purify galectins using three different methods of affinity purification, i.e., lactosyl-Sepharose chromatography for fungal galectinCoprinopsis cinereagalectin 2 (CGL2), nickel-chromatography for histidine-tagged human galectin-7, and glutathione-Sepharose chromatography for GlutathioneS-transferase-tagged (GST-tagged) human galectin-7. Step-by-step instructions are provided for obtaining the above-mentioned recombinant galectins that retain carbohydrate-binding activity and are suitable for conducting biochemical experiments.