FIG. 5.
BIAcore3000 sensorgrams illustrating dosage-dependent binding of synthetic peptides to 10,000 RU of immobilized Hsp72. Peptides were 15 amino acids long and represented the C terminus of Edmonston MV N protein (N peptide), the N peptide with an asparagine-to-aspartic acid substitution at amino acid 4 from the C terminus (NΔ4D peptide), and the NΔ4D peptide with a leucine-to-proline substitution at amino acid 3 from the C terminus (NΔ3P4D peptide). Peptide-Hsp72 responses were corrected for nonspecific interactions with the sensor surface by subtracting the signal recorded when peptides were passed over sensor flow channels lacking Hsp72. The corrected signal was then expressed as a response differential (Resp. Diff.) in RU. Sensorgrams for the interaction between Hsp72 and N peptide concentrations of 25, 50, 100, and 200 μM allowed high-quality global fitting to a 1:1 Langmuir binding model. Calculated equilibrium dissociation constants (KD) and rate constants ka and kd are shown. High-quality global fitting for NΔ4D peptide and NΔ3P4D peptide was based upon sensorgams generated with 100, 200, 500, and 1,000 μM peptide concentrations.