Dataset generated using hyperplexing and click chemistry to monitor temporal dynamics of newly synthesized macrophage secretome post infection by mycobacterial strains.
Dataset generated using hyperplexing and click chemistry to monitor temporal dynamics of newly synthesized macrophage secretome post infection by mycobacterial strains.
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DOI：
10.1016/j.dib.2016.08.055
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发表时间：
2016-12
期刊：
影响因子：
1.2
通讯作者：
Rao KV
中科院分区：
文献类型：
--
作者：
Kumar A;Jamwal S;Midha MK;Hamza B;Aggarwal S;Yadav AK;Rao KV
在这里，我们提供了基于SILAC和iTRAQ的超复合结合BONCAT的点击化学的数据，以选择性富集四种不同结核分枝杆菌感染后不同时间点THP1巨噬细胞分泌的新合成蛋白。巨噬细胞感染结核分枝杆菌H37Ra、H37Rv、BND433和JAL2287株。从感染后6小时开始至26小时，每隔4小时观察新合成的分泌宿主蛋白。我们首次将BONCAT与混合了SILAC和iTRAQ的超复用（18路复用）相结合。采用两组三联体SILAC编码结核分枝杆菌H37Ra和H37Rv， BND433和JAL2287，各有对照。BONCAT用于丰富新合成蛋白质的分泌组，6-plex iTRAQ标记用于量化捕获的蛋白质组的时间变化。每组18 plex在4个MS重复中运行，采用2种线性和2种非线性分离模式。这种新型的混合方法，结合了三路SILAC和六路iTRAQ，在单次MS运行中实现了18路定量。Hyperplexing使大规模的时空系统生物学研究成为可能，其中大量的样本可以同时进行定量处理。数据可通过ProteomeXchange获得，标识符为ProteomeXchange: PXD004281。
Here we provide data for SILAC and iTRAQ based hyperplexing combined with BONCAT based click chemistry for selective enrichment of newly synthesized proteins secreted by THP1 macrophages at various time points after infection with four different strains of Mycobacterium tuberculosis. The macrophages were infected with H37Ra, H37Rv, BND433 and JAL2287 strains of M. tuberculosis. Newly-synthesized secreted host proteins were observed, starting from six hours post-infection till 26 h, at 4 h intervals. We have combined BONCAT with hyperplexing (18-plex), which blends SILAC and iTRAQ, for the first time. Two sets of triplex SILAC were used to encode the strains of M. tuberculosis - H37Ra & H37Rv in one and BND433 & JAL2287 in another with a control in each. BONCAT was used to enrich the secretome for newly synthesized proteins while 6-plex iTRAQ labeling was employed to quantify the temporal changes in the captured proteome. Each set of 18-plex was run in 4 MS replicates with two linear and two non-linear separation modes. This new variant of hyperplexing method, combining triplex SILAC with 6-plex iTRAQ, achieves 18-plex quantitation in a single MS run. Hyperplexing enables large scale spatio-temporal systems biology studies where large number of samples can be processed simultaneously and in quantitative manner. Data are available via ProteomeXchange with identifier ProteomeXchange: PXD004281.