Kit receptor dimerization is driven by bivalent binding of stem cell factor.
Most growth factors and cytokines activate their receptors by inducing dimerization upon binding. We have studied binding of the dimeric cytokine stem cell factor (SCF) to the extracellular domain of its receptor Kit, which is a receptor tyrosine kinase similar to the receptors for platelet-derived growth factor and colony-stimulating factor-1. Calorimetric studies show that one SCF dimer binds simultaneously to two molecules of the Kit extracellular domain. Gel filtration and other methods show that this results in Kit dimerization. It has been proposed that SCF-induced Kit dimerization proceeds via a conformational change that exposes a key receptor dimerization site in the fourth of the five immunoglobulin (Ig)-like domains in Kit. We show that a form of Kit containing just the first three Ig domains (Kit-123) binds to SCF with precisely the same thermodynamic parameters as does Kit-12345. Analytical ultracentrifugation, light scattering, and gel filtration show that Kit-123 dimerizes upon SCF binding in a manner indistinguishable from that seen with Kit-12345. These data argue that the fourth Ig-like domain of Kit is not required for SCF-induced receptor dimerization and provide additional support for a model in which bivalent binding of the SCF dimer provides the driving force for Kit dimerization.
Lemmon
M A
MA
Department of Pharmacology, New York University Medical Center, New York, New York 10016, USA.
Pinchasi
D
D
Zhou
M
M
Lax
I
I
Schlessinger
J
J
D002850:Chromatography, Gel; D006801:Humans; D008024:Ligands; D046911:Macromolecular Substances; D011485:Protein Binding; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*; D011994:Recombinant Proteins; D019089:Stem Cell Factor / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014461:Ultracentrifugation
D008024:Ligands; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D019009:Proto-Oncogene Proteins c-kit