Structure of human anti-cancer drug resistance by hABCG2

hABCG2的人类抗癌耐药性结构

• 批准号: 7490682
• 负责人: STEPHEN G ALLER
• 金额: $ 4.2万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2009-07-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7490682
• 关键词: ABCG2 gene; ATP phosphohydrolase; Amino Acids; Antineoplastic Agents; Arts; Binding; Biological; Biological Assay; Cell-Free System; Cells; Characteristics; Chemicals; Chemistry; Class; Crystallization; Crystallography; Detergents; Dimensions; Doctor of Philosophy; Drug Transport; Drug resistance; Effectiveness; Electron Microscopy; Electrons; Electrophoresis; Equipment; Family; Fellowship; Fermentation; Folic Acid Antagonists; Goals; Health; Homidium Bromide; Human; Individual; Integral Membrane Protein; Learning; Link; Lipids; Literature; Malignant Neoplasms; Membrane Proteins; Methotrexate; Motivation; Mutation; Names; Negative Staining; Orthologous Gene; Output; P-Glycoproteins; Pharmaceutical Preparations; Pharmacologic Substance; Phospholipids; Pichia; Preparation; Production; Protein Family; Proteins; Protocols documentation; Publishing; Pump; Relapse; Resistance; Resolution; Rhodamine; Rhodamines; Robotics; Role; Screening procedure; Solutions; Structure; Synchrotrons; System; Techniques; Training; Vesicle; Work; Yeasts; chemotherapy; design; dimer; experience; glycosylation; human ABCG2 protein; inhibitor/antagonist; interest; member; milligram; neoplastic cell; next generation; reconstitution; success; three dimensional structure; tool; two-dimensional

DESCRIPTION (provided by applicant): Spontaneous mutations of the human ATP-binding cassette breast cancer resistance protein, BCRP (ABCG2), convey tumor cell resistance by exporting classical anti-cancer agents, often resulting in a fatal relapse of chemotherapy. Human ABCG2 is a homodimeric integral membrane protein with a unique and compact "half-transporter" design and does not require any glycosylation to function as a drug export pump. These characteristics may make this protein the best candidate among human multidrug export pumps for high-resolution structure determination. The protocols for the expression of low milligram quantities of ABCG2 from the yeast Pichia pastoris have already been developed. Production of even higher quantities of protein from cell-free systems will be optimized now that non-ionic detergents that stabilize hABCG2 have already been identified. The goal of this proposal is to solve the structure of human ABCG2 by x-ray crystallography to visualize the mechanisms by which this clinically important protein drives anti-cancer pharmaceuticals out of cells. "Click" chemistry techniques will be applied to purified ABCG2 protein and protein crystals to identify building blocks of specific inhibitors to this drug export pump. A high-resolution crystal structure will be used in combination with the output from the "click" chemistry to enable the design of next-generation anti-cancer pharmaceuticals that will specifically block ABCG2 activity and increase the effectiveness of chemotherapy in several human cancers.

描述（由申请方提供）：人ATP结合盒乳腺癌耐药蛋白BCRP（ABCG 2）的自发突变通过输出经典抗癌药物传递肿瘤细胞耐药性，通常导致化疗的致命复发。人ABCG 2是具有独特和紧凑的“半转运蛋白”设计的同源二聚体整合膜蛋白，并且不需要任何糖基化来作为药物输出泵发挥作用。这些特性可能使该蛋白质成为人类多药输出泵中用于高分辨率结构测定的最佳候选者。已经开发了从酵母毕赤酵母表达低毫克量ABCG 2的方案。由于已经鉴定出了稳定hABCG 2的非离子去污剂，因此将优化无细胞系统中更大量的蛋白质的生产。该提案的目标是通过X射线晶体学解决人类ABCG 2的结构，以可视化这种临床重要蛋白质将抗癌药物从细胞中排出的机制。“点击”化学技术将应用于纯化的ABCG 2蛋白和蛋白晶体，以鉴定该药物输出泵的特异性抑制剂的构建块。高分辨率的晶体结构将与“点击”化学的输出相结合，以设计下一代抗癌药物，这些药物将特异性地阻断ABCG 2活性，并增加化疗在几种人类癌症中的有效性。