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. 2009 Jul 21;48(35):8422–8434. doi: 10.1021/bi900606y

Figure 2.

Figure 2

Construction of Slc11a2 proteins containing hemagglutinin (HA) epitope tags. The sites for insertion of YPYDVPDYA hemagglutinin (HA) epitopes in individual Slc11a2 proteins are indicated, along with the numerical designation of the construct (NT, 1−11, CT). These have been superimposed on the hydropathy plot of the Slc11a2 protein produced by the TOPPRED software package using a 20-residue long sliding window (core window, 12 residues; wedge windows, 4 residues). The default parameters suggested for predicting the presence of TMDs in eukaryotic proteins were used, such as the Kyte−Doolittle cutoff values (lower, 0.6; putative; upper, 1.0; certain). Orange arrowheads indicate insertion sites that produced functional proteins in which the polarity of the tag could be established by immunofluorescence. Black arrowheads indicate insertion sites that caused partial or complete loss of transport. Single arrowheads represent single epitope tags while double arrowheads (5b, 8b, 11b) represent constructs with two epitope tags inserted side by side. Boxes at the top of the graph indicate the location of the proposed 2-fold structural symmetry in Slc11a2.