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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: Biomaterials. 2016 Aug 4;105:222–227. doi: 10.1016/j.biomaterials.2016.08.002

Figure 3.

Figure 3

Ni-NTA-nanogold binding assay for determining the orientation of C-His SPP1 connector in the lipid membrane with 92% of the connectors with its N terminal outside the liposome. (A) Schematic. (B) Representative current trace showing TAT peptide translocation (300 nM) through a single connector and subsequent binding of Ni-NTA-nanogold (1.8 nm; 300 pM added into trans-chamber) to C-terminal His-tag of connector. (C) Quantification of the orientation adopted by connectors from 312 insertion events. Cis-chamber: 1 M KCl, 5 mM HEPES, pH 7.9; trans-chamber: 0.15 M KCl, 5 mM HEPES, pH 7.9. Applied voltage: −50 mV.