Bajinting Adam, Ng Ho Leung
Department of Chemistry, University of Hawaii at Manoa, Honolulu, HI, United States of America.
School of Medicine, St. Louis University, St. Louis, MO, United States of America.
PeerJ. 2017 Jun 29;5:e3512. doi: 10.7717/peerj.3512. eCollection 2017.
Fibroblast growth factor receptors (FGFRs) are a family of receptor tyrosine kinases containing three domains: an extracellular receptor domain, a single transmembrane helix, and an intracellular tyrosine kinase domain. FGFRs are activated by fibroblast growth factors (FGFs) as part of complex signal transduction cascades regulating angiogenesis, skeletal formation, cell differentiation, proliferation, cell survival, and cancer. We have developed the first recombinant expression system in to produce a construct of human FGFR2 containing its transmembrane and extracellular receptor domains. We demonstrate that the expressed construct is functional in binding heparin and dimerizing. Size exclusion chromatography demonstrates that the purified FGFR2 does not form a complex with FGF1 or adopts an inactive dimer conformation. Progress towards the successful recombinant production of intact FGFRs will facilitate further biochemical experiments and structure determination that will provide insight into how extracellular FGF binding activates intracellular kinase activity.
成纤维细胞生长因子受体（FGFRs）是一类受体酪氨酸激酶家族，包含三个结构域：一个细胞外受体结构域、一个单一的跨膜螺旋和一个细胞内酪氨酸激酶结构域。FGFRs 被成纤维细胞生长因子（FGFs）激活，作为调节血管生成、骨骼形成、细胞分化、增殖、细胞存活和癌症的复杂信号转导级联反应的一部分。我们已开发出首个重组表达系统，用于生产包含其跨膜和细胞外受体结构域的人 FGFR2 构建体。我们证明所表达的构建体在结合肝素和二聚化方面具有功能。尺寸排阻色谱法表明，纯化的 FGFR2 不与 FGF1 形成复合物，也不采用无活性的二聚体构象。完整 FGFRs 的成功重组生产方面的进展将有助于进一步的生化实验和结构测定，这将深入了解细胞外 FGF 结合如何激活细胞内激酶活性。
