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. 2014 Jan 6;289(8):5285–5295. doi: 10.1074/jbc.M113.523639

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Ankyrin-B associates with Ca_v2.1 and Ca_v2.2. A, ankyrin-B comprises a membrane-binding domain, spectrin-binding domain, and a C-terminal regulatory domain. B–D, GST-ankyrin-B MBD associates with both Ca_v2.1 and Ca_v2.2 from detergent-soluble lysates of cortex (B), cerebellum (C), and brain stem (D). No interaction was observed between lysates and GST-ankyrin-B SBD, GST-ankyrin-B CTD, or GST alone. E, GST fusion proteins were utilized for pull-down experiments. Fusion proteins were purified from bacteria and analyzed by Coomassie Blue staining. Whereas GST migrates at 29 kDa, GST-ankyrin-B MBD migrates at 94 kDa. GST-ankyrin-B SBD migrates at 62 kDa, and GST-ankyrin-B CTD migrates at 79 kDa.