Studies have shown that depending on the substitution pattern, microtubule (MT)-targeting 1,2,4-triazolo[1,5-a]pyrimidines (TPDs) can produce different cellular responses in mammalian cells that may be due to these compounds interacting with distinct binding sites within the MT structure. Selected TPDs are also potently bioactive against the causative agent of human African trypanosomiasis, Trypanosoma brucei, both in vitro and in vivo. So far, however, there has been no direct evidence of tubulin engagement by these TPDs in T. brucei. Therefore, to enable further investigation of anti-trypanosomal TPDs, a TPD derivative amenable to photoaffinity labeling (PAL) was designed, synthesized, and evaluated in PAL experiments using HEK293 cells and T. brucei. The data arising confirmed specific labeling of T. brucei tubulin. In addition, proteomic data revealed differences in the labeling profiles of tubulin between HEK293 and T. brucei, suggesting structural differences between the TPD binding site(s) in mammalian and trypanosomal tubulin.

Center for Discovery and Innovation in Parasitic Diseases Skaggs School of Pharmacy and Pharmaceutical Sciences University of California San Diego 9500 Gilman Drive La Jolla CA 92093 USA

Center for Neurodegenerative Disease Research Perelman School of Medicine University of Pennsylvania 3600 Spruce Street Philadelphia Pennsylvania 19104 USA

Department of Chemistry and Biochemistry University of California San Diego 9500 Gilman Drive La Jolla CA 92093 USA

School of Pharmacy and Pharmaceutical Sciences Cardiff University Cardiff CF103NB U K

Vysoká Škola Chemicko Technologická Praha Department of Organic Chemistry Technická 5 Prague 16628 Czech Republic

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