QI-FRET: a new tool in Receptor Tyrosine Kinase research

QI-FRET：受体酪氨酸激酶研究的新工具

• 批准号: 8588339
• 负责人: Kalina Hristova
• 金额: $ 32.31万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8588339
• 关键词: Biological; Cell membrane; Cells; Chinese Hamster Ovary Cell; Dimerization; Disease; Energy Transfer; Fibroblast Growth Factor Receptors; Fluorescence; Fluorescence Resonance Energy Transfer; Free Energy; Genes; Goals; Growth Disorders; Heterodimerization; Homo; Human Pathology; Image; Knowledge; Lateral; Length; Ligands; Link; Maps; Measures; Membrane; Methodology; Methods; Microscope; Pathology; Process; Proteins; Receptor Protein-Tyrosine Kinases; Research; Signal Transduction; Skeletal system; Systems Development; Universities; Vesicle; Work; cancer therapy; dimer; human disease; malignant breast neoplasm; novel; overexpression; protein expression; public health relevance; receptor; research study; tool

DESCRIPTION (provided by applicant): QI-FRET: a new tool in Receptor Tyrosine Kinase research Kalina Hristova, Johns Hopkins University Ligand-independent homo and heterodimerization of receptor tyrosine kinases (RTKs) is a process that is not well understood despite being implicated in various human pathologies. The lack of knowledge is mainly due to a lack of an appropriate methodology to measure the dimerization energetics of full-length RTK. The lack of knowledge is, in turn, a bottleneck in developing effective RTK-targeted therapies. Here we propose to (i) develop a quantitative imaging FRET (QI-FRET) method that yields dimerization free energies in plasma membrane-derived vesicles and to (ii) characterize the homo and heterodimerization free energies of the ErbB and FGF receptors, chosen because of their very strong link to human disease. Upon the completion of this work, we will be able to predict the degree of ligand-independent dimerization, and thus biological activity, as a function of RTK expression. The method that will be established and the knowledge gained will ultimately aid the search and refinement of effective RTK-targeted treatments for cancers and growth disorders.

描述(由申请人提供)：QI-FRET：受体酪氨酸激酶研究的新工具卡琳娜·赫里斯托娃，约翰霍普金斯大学配体非依赖性受体酪氨酸激酶(RTK)的同源和异源二聚化是一个尽管与多种人类病理学有关但尚未被很好理解的过程。缺乏知识的主要原因是缺乏适当的方法学来测量全长RTK的二聚化能量学。知识的缺乏反过来又是开发有效的RTK靶向疗法的瓶颈。在这里，我们建议(I)建立一种定量成像FRET(QI-FRET)方法，该方法在质膜衍生的小泡中产生二聚化自由能，并且(Ii)表征ErbB和成纤维细胞生长因子受体的同位和异二聚化自由能，因为它们与人类疾病有非常强的联系。这项工作完成后，我们将能够预测与配体无关的二聚化程度，从而预测作为RTK表达的函数的生物活性。即将建立的方法和获得的知识最终将有助于寻找和改进有效的RTK靶向治疗癌症和生长障碍的方法。

项目成果

Direct assessment of the effect of the Gly380Arg achondroplasia mutation on FGFR3 dimerization using quantitative imaging FRET.

• DOI: 10.1371/journal.pone.0046678
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Placone J;Hristova K
• 通讯作者: Hristova K

Glycophorin A transmembrane domain dimerization in plasma membrane vesicles derived from CHO, HEK 293T, and A431 cells.

源自 CHO、HEK 293T 和 A431 细胞的质膜囊泡中血型糖蛋白 A 跨膜结构域二聚化。

• DOI: 10.1016/j.bbamem.2013.03.022
• 发表时间: 2013
• 期刊: Biochimica et biophysica acta
• 作者: Sarabipour,Sarvenaz;Hristova,Kalina
• 通讯作者: Hristova,Kalina

Highly efficient macromolecule-sized poration of lipid bilayers by a synthetically evolved peptide.

• DOI: 10.1021/ja500462s
• 发表时间: 2014-03-26
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Wiedman G;Fuselier T;He J;Searson PC;Hristova K;Wimley WC
• 通讯作者: Wimley WC

Testing the limits of rational design by engineering pH sensitivity into membrane-active peptides.

• DOI: 10.1016/j.bbamem.2014.12.023
• 发表时间: 2015-04
• 期刊: Biochimica et biophysica acta
• 作者: Wiedman G;Wimley WC;Hristova K
• 通讯作者: Hristova K