XRAY STRUCTURE DETERMINATION OF RETINAL SPECIFIC DEHYDROGENASE

视网膜特异性脱氢酶的 X 射线结构测定

• 批准号: 6667814
• 负责人: MARCIA E. NEWCOMER
• 金额: $ 14.27万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2003-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6667814
• 关键词: bioimaging /biomedical imaging; biological products; biomedical resource; proteins; radiography; structural biology

The fibroblast growth factor (FGF) family is comprised of a group of at least nine cytokines which are characterized in part by binding to heparin, and which share 30-55% identity at the amino acid level. FGFs bind to a series of receptors among which is FGFR1 consisting of an intracellular split tyrosine portion, a transmembrane region, and an extracellular region comprised of two or three Ig domains, D(I), D (II), and D (III). In adult organisms, the FGFs are thought to play a role in would healing, in the pathology of tumor growth, rheumatoid arthritis, diabetic retinopathy, and psoriasis. New antagonists of FGF/FGFR1 interactions are expected to find applications for the control of angiogenesis and disease states that are neovascularization depdendent such as tumor growth and metastasis. To further understand the central role that the D(II) domain plays in the binding mechanism of the FGF system, we have undertaken a protein crystallization and x-ray diffraction study of this 118 residue fragment of FGFR1 expressed in E. coli. While we have obtained x-ray diffraction data to ~3
resolution from ~75 mm sized crystals using a storage phosphor detector and a Cu rotating anode source, we feel the combination of synchrotron radiation and cryocooling will give better quality and higher resolution data on these weakly diffracting crystals of the native protein. Phasing of the reflections will be done by MIR employing low resolution data collected using a rotating anode source.

成纤维细胞生长因子 (FGF) 家族由一组成员组成 至少九种细胞因子，其部分特征在于结合 与肝素相比，它们在氨基酸水平上具有 30-55% 的同一性。 FGF 与一系列受体结合，其中 FGFR1 包括 细胞内分裂酪氨酸部分、跨膜区和 由两个或三个 Ig 结构域组成的胞外区域，D(I)、D (II)和D(III)。 在成体生物体中，FGF 被认为发挥着 在愈合、肿瘤生长、类风湿病理学中的作用 关节炎、糖尿病视网膜病变和牛皮癣。 新的对手 FGF/FGFR1 相互作用有望找到应用 控制血管生成和新血管形成的疾病状态 取决于肿瘤的生长和转移等。 为了进一步了解 D(II) 结构域在结合机制中发挥的核心作用 FGF系统的研究，我们进行了蛋白质结晶和 FGFR1 的 118 个残基片段的 X 射线衍射研究 在大肠杆菌中表达。 虽然我们已经获得了X射线衍射数据 使用存储荧光粉从约 75 毫米大小的晶体获得约 3 的分辨率 探测器和铜旋转阳极源，我们感受到的组合 同步加速器辐射和低温冷却将提供更好的质量和 这些弱衍射晶体的更高分辨率数据 天然蛋白质。 反射的定相将由 MIR 完成 采用使用旋转阳极源收集的低分辨率数据。