Analysis of the ligand-binding domain of macrophage colony-stimulating receptor

RNAs have been extracted from human placenta. Extracellular regions of M-CSFR, D1-3, D2-3 and D3 motifs have been amplified with RT-PCR. The proteins have been expressed inE.Coli. Enzyme-linked immusorbent assay (EIA) shows that recombinant D1-3 possesses binding ability 3 times that of D2-3.Kd, the dissociation constant of the former is 11 nmol/L, and that of latter is 39 nmol/L. D3 lacks binding ability. These data suggest that D2-3 is the main site for M-CSF binding, D1 is an assistant site and also contributes to the conformation of site for ligand binding.