Lemmon M A, Pinchasi D, Zhou M, Lax I, Schlessinger J
Department of Pharmacology, New York University Medical Center, New York, New York 10016, USA.
J Biol Chem. 1997 Mar 7;272(10):6311-7. doi: 10.1074/jbc.272.10.6311.
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.
大多数生长因子和细胞因子通过结合时诱导二聚化来激活其受体。我们研究了二聚体细胞因子干细胞因子(SCF)与其受体Kit的细胞外结构域的结合,Kit是一种受体酪氨酸激酶,类似于血小板衍生生长因子和集落刺激因子-1的受体。量热研究表明,一个SCF二聚体同时与两个Kit细胞外结构域分子结合。凝胶过滤和其他方法表明,这会导致Kit二聚化。有人提出,SCF诱导的Kit二聚化是通过构象变化进行的,该构象变化会暴露Kit五个免疫球蛋白(Ig)样结构域中第四个结构域中的关键受体二聚化位点。我们表明,仅包含前三个Ig结构域的Kit形式(Kit-123)与SCF结合时的热力学参数与Kit-12345完全相同。分析超速离心、光散射和凝胶过滤表明,Kit-123在与SCF结合时二聚化,其方式与Kit-12345所见的方式无法区分。这些数据表明,Kit的第四个Ig样结构域对于SCF诱导的受体二聚化不是必需的,并为SCF二聚体的二价结合为Kit二聚化提供驱动力的模型提供了额外支持。