Fig. 3
From: APC/CCDH1 synchronizes ribose-5-phosphate levels and DNA synthesis to cell cycle progression
TKTL1 depletes TKT by forming stable heterodimers with TKT. a Interacting proteins of TKTL1 and TKT. Tandem affinity purification identification of TKTL1- (left) or TKT-interacting (right) proteins. b TKTL1 binds to TKT. TKTL1 and TKT associations were examined by reciprocal co-immunoprecipitation as indicated. c Affinity purified TKTL1 from ccRCC and co-purification of endogenous TKT. d Binding affinity of TKT to TKTL1 and TKT was analyzed by SPR. The right panel shows the purity of both TKTL1 and TKT. CBB, Coomassie brilliant blue. e Gel filtration assays for determining heterodimers MSH6 (152 kDa) and BSA (66 kDa) proteins were used as molecular weight markers. The right panel shows the purity of both TKTL1 and TKT. f Native gel electrophoresis for detecting formation of the TKT dimer, TKTL1-TKT heterodimer, and TKTL1 monomer. g, h (g) TKT was co-expressed with sequentially truncated TKTL1 species and (h) TKTL1 was co-expressed with sequentially truncated TKT species in HEK293T cells; interactions between the overexpressed proteins were analyzed by co-immunoprecipitation. Binding (+) and no binding (−) between species are indicated. i, j (i) TKTL1 without the central portion was co-expressed with TKT and (j) TKT without the central portion was co-expressed with TKTL1; interactions between the overexpressed proteins were analyzed by co-immunoprecipitation. Full-length blots are presented in Supplementary Fig. 10
