Proteomics analysis of lysine methylated proteins from Leishmania donovani strain 1S: wild type and set mutants

This study was performed with the goal of understanding the roles of Leishmania donovani SET7 and SET29 proteins. Using homologous recombination, set7 and set29 genes were knocked out, thereby creating set7 and set29 mutant parasites. Whole cell extracts were isolated from wild type parasites, set7 mutant parasites and set29 mutant parasites, followed by immunoprecipitation of lysine methylated proteins using anti-methyllysine antibody. The immunoprecipitated proteins were identified by LC-MS analysis.

本研究旨在阐明杜氏利什曼原虫（Leishmania donovani）SET7与SET29蛋白的生物学功能。本研究通过同源重组技术对set7和set29基因进行敲除，成功构建set7与set29突变寄生虫株。分别从野生型寄生虫株、set7突变寄生虫株及set29突变寄生虫株中提取全细胞提取物，随后利用抗甲基赖氨酸抗体对赖氨酸甲基化蛋白进行免疫沉淀富集。最终通过液相色谱-质谱联用（LC-MS）分析对免疫沉淀得到的靶蛋白进行鉴定。