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首页> 外文期刊>Protein Expression and Purification >Purification and characterization of the acyltransferase involved in biosynthesis of the major mycobacterial cell envelope glycolipid - Monoacylated phosphatidylinositol dimannoside
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Purification and characterization of the acyltransferase involved in biosynthesis of the major mycobacterial cell envelope glycolipid - Monoacylated phosphatidylinositol dimannoside

机译：参与主要分枝杆菌细胞膜糖脂生物合成的酰基转移酶的纯化和表征-单酰化磷脂酰肌醇二甘露糖苷

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摘要

Phosphatidylinositol mannosides are essential structural components of the mycobacterial cell envelope. They are implicated in host-pathogen interactions during infection and serve as a basis for biosynthesis of other unique molecules with immunomodulatory properties - mycobacterial lipopolysaccharides lipoarabinomannan and lipomannan. Acyltransferase Rv2611 is involved in one of the initial steps in the assembly of these molecules in Mycobacterium tuberculosis - the attachment of an acyl group to position-6 of the 2-linked mannosyl residue of the phosphatidylinositol mannoside anchor. Although the function of this enzyme was annotated 10 years ago, it has never been completely biochemically characterized due to lack of the pure protein. We have successfully overexpressed and purified MSMEG_2934, the ortholog of Rv2611c from the non-pathogenic model organism Mycobacterium smegmatis mc~2155 using mycobacterial pJAM2 expression system, which allowed confirmation of its in vitro acyltransferase activity, and establishment of its substrate specificity.
机译：磷脂酰肌醇甘露糖苷是分枝杆菌细胞包膜的基本结构成分。它们与感染过程中的宿主-病原体相互作用有关，并作为生物合成具有免疫调节特性的其他独特分子的基础-分枝杆菌脂多糖lipoarabinomannan和lipomannan。酰基转移酶Rv2611参与结核分枝杆菌中这些分子的组装的初始步骤之一-酰基与磷脂酰肌醇甘露糖苷锚定的2-连接的甘露糖基残基的6位连接。尽管该酶的功能在10年前就已注释，但由于缺乏纯蛋白质，从未对其进行完全的生化鉴定。我们已经成功地使用分枝杆菌pJAM2表达系统从非致病性模型生物耻垢分枝杆菌mc〜2155中过表达和纯化了Rs2611c的直系同源基因MSMEG_2934，这可以确认其体外酰基转移酶活性，并确定其底物特异性。

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