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. Author manuscript; available in PMC: 2006 Jan 26.
Published in final edited form as: Biochemistry. 2004 Sep 7;43(35):11175–11186. doi: 10.1021/bi048848d

Figure 8.

Figure 8

Interactions of ALG-2 and mutants with Alix/AIP1. In panel A, ALG-2 and dF122 (400 nM each) were injected over the immobilized Alix/AIP1 surface in the presence of 50 μM CaCl2 using the COINJECT program outlined in the Materials and Methods. In panel B, similarly, ALG-2 and the three EF-hand mutants, E47D, E114D, and D169A were compared using the COINJECT method. In panel C, ALG-2 and dNALG-2 (500 nM each) in the presence of 50 μM CaCl2 with or without 200 μM decapeptide were analyzed by surface plasmon resonance for interaction with immobilized Alix/AIP1 using the KINJECT program. Flat plateaus as seen with KINJECT were not seen with COINJECT due to methodological differences between the two programs; however, slower dissociation rates were observable with COINJECT by retaining Ca2+ concentration in the dissociation phase. Experiments were conducted at least twice using duplicate samples in each experiment.