Crystal structures of IFT70/52 and IFT52/46 provide insight into intraflagellar transport B core complex assembly

Shown is a Coomassie stained gel after SDS-PAGE analysis of a pull-down experiment of His-tagged IFT70 or SUMO-tagged IFT88 with 3 different truncated GST-tagged IFT52 constructs.This experiment revealed that GST-tagged IFT52(330-381) pulls down IFT70, whereas GST-tagged IFT52(281-329) binds to IFT88. (left to right) Lane 1: molecular weight marker. Lanes 2 to 7: total bacterial lysates. Lane 2: co-expression of GST-tagged IFT52(281-381) and His-tagged IFT70. Lane 3: co-expression of GST-tagged IFT52(281-329) and His-tagged IFT70. Lane 4: co-expression of GST-tagged IFT52(330-381) and His-tagged IFT70. Lane 5: co-expression of GST-tagged IFT52(281-381) and SUMO-tagged IFT88. Lane 6: co-expression of GST-tagged IFT52(281-329) and SUMO-tagged IFT88. Lane 7: co-expression of GST-tagged IFT52(330-381) and SUMO-tagged IFT88. Lanes 8 to 13: Samples were loaded after GST pull-down of GST-tagged truncated IFT52 constructs co-expressed with His-tagged IFT70 or SUMO-tagged IFT88. The constructs were loaded in the same order as for lanes 2 to 7. The prominent bands correspond to: Lane 8: His-tagged IFT70 (upper band) and GST-tagged IFT52(281-381) (lower band). Lane 9: GST-tagged IFT52(281-329) (upper band) and GST (lower band). Lane 10: His-tagged IFT70 (upper band) and GST-tagged IFT52(330-381) (lower band). Lane 11: SUMO-tagged IFT88 (upper band) and GST-tagged IFT52(281-381) (lower band). Lane 12: SUMO-tagged IFT88 (upper band), GST-tagged IFT52(281-329) (middle band), and GST (lowest band). Lane 13: GST-tagged IFT52(330-381) (upper band), GST-tagged IFT52(281-329) (middle band), and GST (lower band).