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. Author manuscript; available in PMC: 2012 Jul 19.
Published in final edited form as: Biochemistry. 2011 Jun 24;50(28):6208–6216. doi: 10.1021/bi200508f

Figure 4.

Figure 4

LRP-CT microdomains binding to (A) Fe65 WW-PID1-PID2 236-662, (B) Fe65 WW-PID1 236-512, or (C) Fe65 PID2 536-662. Microdomains containing Y4473 (Cys-amino aminohexanoic acid-VEIGNPTY4473KMYEGGE) or Y4507 (Cys-amino aminohexanoic acid-TNFTNPVY4507ATLY) in both tryrosine-phosphorylated and non-phosphorylated forms were immobilized using sulfolink coupling gel. Each microdomain (60 μg) was incubated with 180 nM –400 nM Fe65 for 1 hour at 4°C. Sulfolink resin alone was used as a negative control. Bound Fe65 was visualized by western blotting with anti-Fe65 antibody (Millipore, 3H6) or anti-HIS antibody (Quiagen, Penta-His Ab). Fe65 WW-PID1-PID2 bound to both the phospho- and non-phospho-forms of the Y4507-containing microdomain of LRP. Panel B shows that only the phospho-form binds to Fe65 PID1 and Panel C shows that only the non-phospho-form binds to Fe65 PID2.