Structure and Function of Concentrative Nucleoside Transporters

浓缩核苷转运蛋白的结构和功能

基本信息

批准号：
9024572
负责人：
Seok-Yong Lee
金额：
$ 29.41万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9024572
关键词：
Adenosine Adopted Adverse effects Amino Acids Antineoplastic Agents Antiviral Agents Ara-C Architecture Binding Binding Sites Biological Assay Biological Availability Biological Models Cancer cell line Cell membrane Cells Chemicals Complex Coupled Coupling DNA Data Detergents Development Drug Targeting Drug Transport Electrodes Family Goals Health Homologous Gene Human Human body Integral Membrane Protein Intestines Ions Kidney Lighting Liver Membrane Molecular Molecular Conformation Mutation Nucleic Acids Nucleoside Transporter Nucleosides Organ Organism Pharmaceutical Preparations Pharmacology Pharmacotherapy Physiological Physiology Positioning Attribute Property Protein Engineering RNA chemical synthesis Radioactive Reagent Resistance Ribavirin Role Sequence Homology Signal Transduction Sodium Source Specificity Structure Substrate Specificity Testing Uridine Vibrio cholerae Zidovudine base cancer cell cancer type cytotoxic design gemcitabine improved insight mutant novel therapeutics protein transport research study response screening solute stoichiometry voltage clamp

项目摘要

DESCRIPTION (provided by applicant): Nucleoside transporters (NTs) are integral membrane proteins that are designed to transport nucleosides into the human body. Nucleoside transport across cell membranes is physiologically important as it provides the major source of nucleosides for DNA/RNA synthesis in cells and it is responsible for the termination of adenosine signaling. Another important fact about NTs is that many nucleoside-derived anticancer and antiviral drugs are transported into cells by way of NTs. Despite their importance in physiology and pharmacology, the molecular mechanism of NTs is largely unknown due to the lack of atomic structures of the transporters. Great advances in the field depend on the structure determination and illumination of the transport mechanism of NTs. Our long- term goal is to understand the fundamental mechanism of selective nucleoside transport by concentrative nucleoside transporters (CNTs) with structural and functional studies. CNTs belong to a family of the solute carrier transporter superfamily (SLC28) and use an ion gradient to transport nucleosides actively against their chemical gradients. We have chosen to use a CNT homolog from Vibrio cholerae (vcCNT) as a model system for our structural and functional studies. vcCNT is an excellent model system to study hCNTs that shares significant sequence homology and functional properties with hCNTs. We solved the crystal structure of vcCNT in complex with uridine. The structure reveals the overall architecture and the nucleoside-binding site of the transporter, providing mechanistic insights of selective nucleoside transport by vcCNT and CNT in general. These new results have led us to propose a combined structure-function study with the following three aims: 1) Determination of the crystal structures of vcCNT in complex with different nucleosides and nucleoside drugs, 2) Functional characterization and comparison of vcCNT with hCNTs, and 3) Structure determination of vcCNT in alternate conformations. The proposed studies will significantly advance our understanding of the mechanism of nucleoside and nucleoside drug transport by prokaryotic and eukaryotic CNTs, which eventually will facilitate the development of new therapeutic reagents.

描述（由申请人提供）：核苷转运蛋白（NT）是一种完整的膜蛋白，旨在将核苷转运到人体中。核苷跨细胞膜转运具有重要的生理意义，因为它为细胞中 DNA/RNA 合成提供核苷的主要来源，并且负责终止腺苷信号传导。关于 NT 的另一个重要事实是，许多核苷衍生的抗癌和抗病毒药物都是通过 NT 转运到细胞中的。尽管 NT 在生理学和药理学中具有重要意义，但由于缺乏转运蛋白的原子结构，其分子机制在很大程度上仍不清楚。该领域的巨大进步取决于 NT 运输机制的结构确定和阐明。我们的长期目标是通过结构和功能研究来了解集中核苷转运蛋白（CNT）选择性核苷转运的基本机制。 CNT 属于溶质载流子转运蛋白超家族 (SLC28)，并使用离子梯度逆其化学梯度主动转运核苷。我们选择使用霍乱弧菌 (vcCNT) 的 CNT 同系物作为我们结构和功能研究的模型系统。 vcCNT 是研究 hCNT 的优秀模型系统，与 hCNT 具有显着的序列同源性和功能特性。我们解析了vcCNT与尿苷复合物的晶体结构。该结构揭示了转运蛋白的整体结构和核苷结合位点，提供了 vcCNT 和 CNT 选择性核苷转运的机制见解。这些新结果使我们提出了一项结构-功能相结合的研究，其目标如下：1) 确定 vcCNT 与不同核苷和核苷药物复合物的晶体结构，2) vcCNT 与 hCNT 的功能表征和比较，以及3）替代构象中vcCNT的结构测定。拟议的研究将显着增进我们对原核和真核碳纳米管核苷和核苷药物转运机制的理解，最终将促进新治疗试剂的开发。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Liposome reconstitution and transport assay for recombinant transporters.

重组转运蛋白的脂质体重建和转运测定。

DOI：
发表时间：
2015
期刊：
影响因子：
0
作者：
Johnson, Zachary Lee;Lee, Seok
通讯作者：
Lee, Seok

Structural basis of nucleoside and nucleoside drug selectivity by concentrative nucleoside transporters.

集中核苷转运蛋白的核苷和核苷药物选择性的结构基础。

DOI：
发表时间：
2014
期刊：
影响因子：
7.7
作者：
Johnson, Zachary Lee;Lee, Jun;Lee, Kiyoun;Lee, Minhee;Kwon, Do;Hong, Jiyong;Lee, Seok
通讯作者：
Lee, Seok

Structural insights into inhibition of lipid I production in bacterial cell wall synthesis.

抑制细菌细胞壁合成中脂质 I 产生的结构见解。

DOI：
发表时间：
2016-05-26
期刊：
影响因子：
64.8
作者：
Chung, Ben C;Mashalidis, Ellene H;Tanino, Tetsuya;Kim, Mijung;Matsuda, Akira;Hong, Jiyong;Ichikawa, Satoshi;Lee, Seok
通讯作者：
Lee, Seok

Crystal structure of a concentrative nucleoside transporter from Vibrio cholerae at 2.4 Å.

霍乱弧菌集中核苷转运蛋白的晶体结构（2.4°）。