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. 2016 Apr 29;2(6):427–441. doi: 10.1021/acsinfecdis.6b00034

Figure 3.

Figure 3

Identification of YnMyr-tagged proteins in (a) BSF and (b) PCF T. brucei. Volcano plots show significance of enrichment over background. Parasites were treated with Myr or YnMyr, lysates labeled by CuAAC, and proteins enriched by biotin–streptavidin interaction and digested by trypsin for LC-MS/MS. Proteins from four (BSF) or six (PCF) replicates (independent sample processing from the lysate stage; includes a biological replicate in each case) were quantified by label-free quantification (MaxLFQ). After filtering to retain only those proteins present in biological duplicate and in 3 of 4 (BSF) or 4 of 6 (PCF) samples, missing intensities were imputed from a normal distribution chosen to mimic noise level, and a modified t test with permutation-based FDR statistics was applied (250 permutations) to compare Myr and YnMyr groups. BSF and PCF: FDR = 0.001; s0 = 1. Proteins containing an N-terminal glycine (MG) and those for which the YnMyr-modified peptide was identified using reagents AzRB or AzRTB in the respective lifestage are indicated. ARF* indicates multiple proteins in this proteinGroup. See also Supporting Information Tables S1–S3.