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Interaction of MKLP1 with Arf6 in vitro.

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Presentation on theme: "Interaction of MKLP1 with Arf6 in vitro."— Presentation transcript:

1 Interaction of MKLP1 with Arf6 in vitro.
Interaction of MKLP1 with Arf6 in vitro. (A) Domain organization and comparison of the structures of the long and short forms of MKLP1. (B) A MKLP1 region encompassing residues 690−807 is responsible for interaction with Arf6. Lysates of HeLa cells expressing C‐terminally HA‐tagged Arf6(Q67L) were pulled down with GST, GST–MKLP1(690–807) (cMKLP1) or GST–MKLP1(690–789) and subjected to immunoblotting with anti‐HA antibody. (C) GTP‐bound Arf6 interacts with MKLP1. Lysates of HeLa cells expressing C‐terminally HA‐tagged Arf6(WT), Arf6(Q67L) or Arf6(T27N) were pulled down with GST or GST–cMKLP1 and subjected to immunoblotting with anti‐HA antibody. (D) MKLP1 interacts with all Arf isoforms examined. Lysates of HeLa cells expressing C‐terminally HA‐tagged Arf1(Q71L), Arf3(Q71L), Arf5(Q71L) or Arf6(Q67L) were pulled down with GST or GST–cMKLP1 and subjected to immunoblotting with anti‐HA antibody. (E) SPR analysis of the GST–cMKLP1 interaction with Arf1, Arf3, Arf5 or Arf6. Steady‐state resonance (Req) levels were plotted against each Arf concentration. Kd values were calculated from the fitted curves. Hisayoshi Makyio et al. EMBO J. 2012;31: © as stated in the article, figure or figure legend

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