FIG 1.

Identification of ZNF658 as a candidate ZTRE-binding protein. (A) Identification by EMSA of a specific band resulting from binding of a protein factor to the ZTRE in the SLC30A5 promoter that was subsequently analyzed by MALDI-TOF. An IRD-labeled probe (50 fmol), corresponding to the region from positions −156 to +46 of the SLC30A5 gene was electrophoresed through a nondenaturing 5% polyacrylamide gel after incubation with protein extract prepared from Caco-2 cells. Competitor oligonucleotide, including the ZTRE (200-fold excess over probe) was included in the binding reaction as indicated. The arrow indicates the position of the band analyzed. (B) Peptides generated by trypsin-mediated hydrolysis of ZNF658 with masses corresponding to fragments detected by MALDI-TOF mass spectrometry in the protein mixture obtained from the band shown in panel A. The highlighted regions of protein sequence indicate the peptides, which cover 168 of the 1,058 amino acids (16%).