Expression cloning of a receptor for human granulocyte-macrophage colony-stimulating factor.

摘要: Two cDNA clones encoding a receptor for human granulocyte-macrophage colony-stimulating factor (hGM-CSF-R) were isolated by expression screening of library made from placental mRNA. Pools recombinant plasmid DNA electroporated into COS cells which then screened their capacity to bind radioiodinated hGM-CSF using sensitive microscopic autoradiographic approach. The cloned GM-CSF-R precursor is 400 amino acid polypeptide (Mr 45,000) with single transmembrane domain, glycosylated extracellular domain and short (54 acids) intracytoplasmic tail. It does not contain tyrosine kinase nor show homology members the immunoglobulin super gene family, but some significant sequence homologies receptors several other haemopoietic growth factors, including those interleukin-6, erythropoietin interleukin-2 (beta-chain) also prolactin receptor. When transfected directed showing class affinity (KD = 2(-8) nM) specificity GM-CSF interleukin-3. Messenger RNA coding this was detected in variety known display binding, cross-linking experiments revealed similar size cells.