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Figure 3 | BMC Pharmacology

Figure 3

From: Identification of specific calcitonin-like receptor residues important for calcitonin gene-related peptide high affinity binding

Figure 3

Competition binding characteristics of CGRP and AcCGRP(19–37) for wild type and mutant CLRs. Increasing amounts of (A) CGRP or (B) AcCGRP(19–37) were used to compete for specific [125I-Tyr]CGRP(8–37) binding sites on crude HEK293T-RAMP1 membrane preparations transiently transfected, individually with wild type (■), L24A (), L34A () or L24A,L34A (x) CLR mutations. Non-linear regression analysis was used to best fit a sigmoidal curve from the data points of each individual competition binding experiment. From this best fit curve the concentration of competing peptide need to displace 50% of specific radioligand binding (IC50) was estimated and used to calculate the equilibrium dissociation constant (Ki) of unlabled peptide for each CLR. The calculated Ki values of the competing peptides for the L24A, L34A, or L24A,L34A CLR mutants were significantly different (P < .05) from the wild type receptor and are displayed in Table 1. The data presented represents the mean ± S.E. for n = 3–5 experiments performed in duplicate.

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