Figure 2
From: Alp7/TACC-Alp14/TOG generates long-lived, fast-growing MTs by an unconventional mechanism
Binding interactions of S. pombe Alp14, Alp7 and tubulin (A) Full length Alp14 with a C-terminal his or Avi tag and Alp7 with an N-terminal avi-tag were expressed in a baculovirus system and purified using tag-affinity chromatography (see Methods). (B–D) 5–40% glycerol gradient centrifugation of purified Alp14, Alp7 and tubulin (see Methods), SDS-PAGE with Coomassie staining. (B) Alp14 alone and with GTP-brain tubulin and GTP and GDP S. pombe α1β tubulin. Peak fractions are contrast-enhanced for clarity. (C) Alp7 is not shifted by adding tubulin. (D) Alp14 is shifted further by GTP-tubulin than by GDP-tubulin. Alp7 is shifted by Alp14 and the complex is further shifted by GTP-tubulin.
