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. 2011 Jan 26;6(1):e16199. doi: 10.1371/journal.pone.0016199

Figure 1. Human frataxin interacts specifically with ISCU, NFS1 and ISD11.

Figure 1

(A) Mass spectrometry analysis of proteins identified by co-IP with FLAG-tagged frataxin from HeLa mitochondrial enriched fractions. Results represent the proteins specifically identified common to two independent experiments with hFXN-FLAG expressing cells compared to non-transfected cells. Peptides were selected with a stringent filter to avoid a maximum of false positives. Coverage represents the percentage of sequence matching with peptides found in the analysis. An example of complete results for one experiment is given in Table S1. (B) IP obtained in (A) were analyzed by Western blot using specific antibodies against frataxin (intermediate and mature), mitochondrial aconitase (mACO), NFS1, ferrochelatase (FECH), MnSOD and ISCU. MnSOD was used as a control to evaluate non-specific binding on the beads. Inputs correspond to 5µg of mitochondrial HeLa extracts. (C) GST pull-down using GST-hFXN (aa 81–210) and HeLa mitochondrial extracts. Eluted fractions were analyzed by Western blot as in (B) or by coomassie blue staining to detect GST and GST-hFXN.