Identification of telomere length regulation protein 2 homolog (TELO2) as a mammalian target of IVM
(A) Chemical structure of avermectins and immobilized dihydroavermectin B1a (IVM B1a) for target identification.
(B) IVM B1a-binding proteins were identified through a pull-down assay with chemically immobilized IVM B1a. HEK293 cell lysates were incubated with the affinity beads in the absence or presence of soluble IVM as a competitor at 33- or 100-folds. The bound fractions were eluted, resolved by SDS-PAGE, and visualized with silver staining. The arrows indicate the bands that were competed out by the competitor.
(C) Mascot analysis of the data was summarized. TELO2 was identified as an IVM B1a-binding protein. Cluster of K22E, keratins, type II cytoskeletal 2 epidermal, 5, 6A, 6B, and 75; VIME, vimentin; TTI1, TELO2-interacting protein 1 homolog; K2C1, keratin, type II cytoskeletal 1; IPO11, importin-11; SGPL1, sphingosine-1-phosphate lyase 1; K1C9, keratin, type I cytoskeletal 9; cluster of TBB4B, cluster of tubulin beta, beta-2A, beta-4B, and beta-6 chains; K1C10, keratin, type I cytoskeletal 10. See also Data S1.
(D) Binding of TELO2 to the immobilized IVM B1a was confirmed through western blotting with an anti-TELO2 antibody. Free IVM was added as the competitor.