Journal: Pflugers Archiv
Article Title: Interactions between the C-terminus of Kv1.5 and Kv? regulate pyridine nucleotide-dependent changes in channel gating
Figure Lengend Snippet: Binding of the C-terminal domain of Kv to Kvβ (a) Western blots of Kvβ2 (upper panel) and Kvβ3 (middle panel) pulled down by the GST-Kv1.5 C-terminus fusion peptides. Fusion proteins containing 60, 38, or 19 terminal amino acid peptides from Kvα1.5 C-terminus attached to GST or GST with unrelated peptide (Control; 30μ g each) were incubated with lysate of Kvβ2 or Kvβ3 -expressing E.coli (350 μ g total protein). Protein complexes were pulled down using GST·Bind beads, washed and eluted with 10mM glutathione. The eluate was separated by SDS-PAGE and probed with anti-pan-Kvβ antibody, an antibody directed against the C-terminus of Kv1.5 (bait) or GST; (b) Densitometric analysis of the bands in panel a. The density of the Kvβ band precipitated with GST-C60 was assigned a 100% value. †, P
Article Snippet: To further examine the propensity of the C-terminal domain of Kv1.5 to bind ancillary subunits, we compared the amino acid sequence of Kv1.5 C-terminus to that of Kv1.2 and the Drosophila Shaker channel ( ).
Techniques: Binding Assay, Western Blot, Incubation, Expressing, SDS Page