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

Figure 1.

Figure 1

YnMyr labels proteins in T. brucei PCF parasites. (a) Overview of tagging strategy. YnMyr or myristic acid (Myr) was added to T. brucei parasite cultures and incorporated metabolically into lipidated proteins. The alkyne tag was reacted by CuAAC with fluorophore and/or biotin-functionalized azide capture reagents (Supporting Information Figure S1) to allow downstream enrichment and analysis by proteomics and SDS-PAGE. (b) Labeling with YnMyr or Myr (−) in PCF parasites. After 18 h of incubation with probes at 100 μM, parasites were lysed, and proteins were reacted with AzTB and separated by gel for fluorescence scanning. Samples were treated with NaOH or precipitated with chloroform/methanol (C/M) as indicated. (c) Specific PCF labeling is resistant to treatment with Pronase. Coomassie gels are shown in Supporting Information Figure S2.