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. 2014 Nov 25;290(2):1106–1118. doi: 10.1074/jbc.M114.615559

FIGURE 4.

FIGURE 4.

The Cdc34 acidic tail forms cross-links to multiple residues in the Cul1 basic canyon. A, Cul1 Western blot of reactions between either Cdc34 1C or Cdc34 2C proteins and K679C Cul1-Rbx1 in the presence of BMOE. Note that very little product is formed between Cdc34 1C and Cul1, whereas almost all Cul1 is cross-linked to the Cdc34 2C proteins. B, Cdc34 Western blot of the identical reactions shown in A. C, color overlay of the Cul1 (red) and Cdc34 (yellow) Western blots, demonstrating that cross-linked products consist of both Cdc34 and Cul1. D, Cul1 Western blot of reactions between either Cdc34 1C or Cdc34 2C proteins and WT Cul1-Rbx1. E, graph showing the fraction of either WT or K679C Cul1 subunits that cross-linked to either Cdc34 1C or Cdc34 2C proteins. Error bars, S.E. of measurement from duplicate data points. F, color overlay of reactions between WT (red) or K679C (yellow) Cul1-Rbx1 and Cdc34 1C or Cdc34 2C proteins. Note the difference in electrophoretic mobilities for WT Cul1-Cdc34 and K679C Cul1-Cdc34 products.