S100A12的表达、纯化及抗菌活性
Expression, Purification, and Antimicrobial Activity of S100A12.
作者信息
Jackson Emmanuel, Little Saffron, Franklin Dana S, Gaddy Jennifer A, Damo Steven M
机构信息
Department of Life and Physical Sciences, Fisk University.
Tennessee Valley Healthcare Systems, U. S. Dept. of Veterans Affairs; Department of Medicine - Division of Infectious Diseases, Vanderbilt University Medical School;
出版信息
J Vis Exp. 2017 May 13(123):55557. doi: 10.3791/55557.
Calgranulin proteins are important mediators of innate immunity and are members of the S100 class of the EF-hand family of calcium binding proteins. Some S100 proteins have the capacity to bind transition metals with high affinity and effectively sequester them away from invading microbial pathogens in a process that is termed "nutritional immunity". S100A12 (EN-RAGE) binds both zinc and copper and is highly abundant in innate immune cells such as macrophages and neutrophils. We report a refined method for the expression, enrichment and purification of S100A12 in its active, metal-binding configuration. Utilization of this protein in bacterial growth and viability analyses reveals that S100A12 has antimicrobial activity against the bacterial pathogen, Helicobacter pylori. The antimicrobial activity is predicated on the zinc-binding activity of S100A12, which chelates nutrient zinc, thereby starving H. pylori which requires zinc for growth and proliferation.
钙粒蛋白是先天性免疫的重要介质，属于EF手型钙结合蛋白家族的S100类成员。一些S100蛋白能够以高亲和力结合过渡金属，并在一个被称为“营养免疫”的过程中有效地将它们与入侵的微生物病原体隔离开来。S100A12（EN-RAGE）能结合锌和铜，在巨噬细胞和中性粒细胞等先天性免疫细胞中含量很高。我们报告了一种在其活性金属结合构型下表达、富集和纯化S100A12的优化方法。利用这种蛋白质进行细菌生长和活力分析表明，S100A12对细菌病原体幽门螺杆菌具有抗菌活性。这种抗菌活性取决于S100A12的锌结合活性，它螯合营养锌，从而使需要锌进行生长和增殖的幽门螺杆菌饥饿。
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