Fig. 1: PEX39 interacts with members of the import pathway of PTS2-containing proteins.
From: PEX39 facilitates the peroxisomal import of PTS2-containing proteins
a, Depiction of protein–protein interactions for Yjr012c and C6ORF226, per BioGRID25 and BioPlex26,27, respectively. The circles represent proteins and the arrows point from purified proteins to interactors. For Yjr012c, only interactions found in at least two independent studies were considered. For C6ORF226, the combined interactome from HCT116 and HEK293T cells is shown. b, Domain analysis and alignment of Yjr012c and C6ORF226. Probabilities of disorder were determined using IUPred2A56. c, Quantitative proteomic analysis of Pex18 complexes that were affinity purified from soluble fractions of wild-type and Pex18-TPA-expressing yeast grown in oleic acid medium (n = 3). The enrichment of proteins in Pex18 complexes and Q values were determined using the rank-sum method57. Peroxins and PTS2-containing proteins are labelled and/or marked by black dots. The dashed lines indicate a Q-value threshold of 0.05 and a fold-enrichment of 64. d, Immunoblots of anti-FLAG immunoprecipitates prepared from HCT116 cells expressing the indicated proteins. HA denotes the detection of HA-tagged proteins. The dashed lines indicate where different lanes of the same membrane were brought together. For the PHYH, ACAA1 and AGPS blots, solid and open red arrowheads indicate mature and precursor forms of these proteins, respectively. An asterisk indicates a non-specific band. The control protein β-actin (ACTB) should be absent from immunoprecipitates. e, Immunoblots of anti-FLAG immunoprecipitates prepared from HEK293T cells expressing the indicated proteins. The control protein calnexin (CANX) should be absent from immunoprecipitates. The annotation of the immunoblots is otherwise the same as for d. f, Immunoblots of anti-FLAG immunoprecipitates prepared from HEK293T cells expressing FLAG-HA-eGFP or HsPEX39-FLAG-HA. An asterisk indicates a non-specific band. g, Native PAGE and autoradiography of [35S]H6PEX7 pre-incubated with the recombinant proteins GST-HsPEX39, H6PHYH and H6PEX5(1–324), as indicated. The in-gel positions of PEX7 alone, lysate haemoglobin and the complexes PEX7–HsPEX39 (hashtag), PEX7–PHYH–HsPEX39 (ampersand) and PEX7–PEX5–PHYH–HsPEX39 (dollar sign) are indicated. The autoradiograph and corresponding Ponceau S-stained membrane are shown. α, α-helix; IP, immunoprecipitate.
