(a) Schematic representation of the four human prenyltransferases comprised by combinations of α [PTAR1, FNTA (PTAR2), and RabGGTA (PTAR3)] and β (FNTB, PGGT1B and, RabGGTB) subunits, and their substrates. As shown in this study, PTAR1 and RabGGTB interact to form a prenyltransferase that we named GGTase3.
(b) HEK-293T cells were transfected with the indicated GFP-tagged substrates of prenyltransferases for immunoprecipitations and immunoblotting.
(c,d) HEK-293T cells were transfected with the indicated plasmids for immunoprecipitations and immunoblotting. WCE: Whole cell extract; EV: empty vector.
(e) HEK-293T cells were co-transfected with FLAG-tagged PTAR1 and GFP-tagged FBXL2 as indicated. Immunoprecipitations were carried out sequentially using first an anti-FLAG antibody and then an anti-GFP antibody as described in methods. The first elution was done with a FLAG peptide and the second with 1%SDS. The two eluates were then immunoblotted as indicated.