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Fig. 5 | BMC Biochemistry

Fig. 5

From: Antiproliferative factor (APF) binds specifically to sites within the cytoskeleton-associated protein 4 (CKAP4) extracellular domain

Fig. 5

Binding kinetics of as-APF interaction with CKAP4 extracellular domain deletion mutants. Three CKAP4 ED deletion mutants were immobilized onto the Fc2 channel of separate CM5 chips, using amine-coupling, as the ligands; the Fc1 channels were treated similarly but without the protein, serving as reference. Prior to the kinetics assay, binding of each mutant to the APF control peptide was tested, and no specific binding was detected. Multi-cycle kinetics assays were then performed by the injection of various concentrations of as-APF (2.5–40 μM, colored lines) over the CM5 chip surfaces on which CKAP4127–360 (a), CKAP4361–524 (b), and CKAP4525–602 (c) were immobilized via amine-coupling. The adjusted sensorgrams (Fc2-Fc1) were overlaid to calculate binding kinetics. The data were fit using Biacore Evaluation software, and the fitting curves (black lines) as well as the parameters of binding kinetics are shown where applicable. The immobilized RU for each ligand is: CKAP4127–360 at 352 RU, CKAP4361–524 at 585 RU, and CKAP4525–602 at 1914 RU. The heterogeneous ligand model is the best fit for 5A (Chi2 = 0.893); the 1:1 binding model is the best fit for 5B (Chi2 = 0.874); and the two-state binding model is the best fit for 5C (Chi2 = 0.446)

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