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Substrate Specificity Determinants in Nutrient Solute Carrier Transporters

营养溶质载体转运蛋白的底物特异性决定因素

基本信息

批准号：
10735432
负责人：
Avner Schlessinger
金额：
$ 43.78万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-10 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10735432
关键词：
Alanine Allosteric Site Amino Acid Transporter Amino Acids Antiviral Agents Autoimmune Diseases Basic Amino Acid Transport Systems Binding Binding Sites Biochemical Biochemical Pathway Biological Biological Assay Biology Biomass Biophysics Cancer cell line Cell Death Cell Differentiation process Cell Line Cell Proliferation Cells Cellular Metabolic Process Chemicals Collaborations Computing Methodologies Coupled Cryoelectron Microscopy Cysteine Data Development Diabetes Mellitus Disease Docking Drug Delivery Systems Drug Design Drug Targeting Electrophysiology (science)Electrostatics Family Free Energy Future Glutamine Goals Grant Health Human Hydrophobicity Hyperactivity Ion Transport Ions Kinetics Knowledge Laboratories Leucine Libraries Ligands Malignant Neoplasms Malignant neoplasm of prostate Mediating Membrane Membrane Transport Proteins Metabolic Diseases Metabolism Methods Modeling Molecular Molecular Conformation Myocardial Ischemia Neurotransmitters Nutrient Organic Synthesis Pathology Pharmaceutical Preparations Pharmacology Physiological Play Prodrugs Proliferating Proteins Role Serine Shapes Signaling Molecule Site-Directed Mutagenesis Specificity Starvation Structural Models Structure Structure-Activity Relationship Substrate Specificity Sugar Acids Synthesis Chemistry T-Lymphocyte Testing Toxin Work amino acid metabolism analog biophysical techniques cancer cell cancer type computational chemistry design drug discovery inhibitor innovation melanoma neoplastic cell nervous system disorder novel novel strategies rapid growth rational design small molecule solute structural biology therapeutic target tool triple-negative invasive breast carcinoma uptake virtual

项目摘要

SUMMARY Alterations in cell metabolism support rapid growth and proliferation of cells in pathologies such as cancer, au- toimmune disease, and heart ischemia, resulting in increased reliance on the metabolism of amino acids such as glutamine and leucine. Nutrient Solute Carrier (SLC) transporters play a major role in reprogrammed meta- bolic networks by supplying cells with nutrients that are used to build biomass, serve as signaling molecules that enhance cell proliferation and differentiation, or regulate cell death. Our broad goal is to describe the sub- strate and inhibitor specificity determinants in disease-related nutrient SLC transporters and develop unique strategies to modulate their functions. We take an integrative approach that includes computational chemistry methods, coupled with biochemical and biophysical approaches and disease-related cell lines, to char- acterize two amino acid transporters that play a key role in metabolism of rapidly-growing cells: the Alanine- Serine-Cysteine Transporter (SLC1A5, ASCT2), a Na+-dependent amino acid exchanger that modulates intra- cellular glutamine levels, and the Amino Acid Transporter B0+ (ATB0+, SLC6A14), a neutral and cationic amino acid transporter, driven by Na+ and Cl- co-transport. In Aim 1 of this project, we will continue characterizing ASCT2, a well-validated drug target for various patholo- gies (eg triple negative breast cancer and prostate cancer). Despite recent advancements in our understanding of ASCT2 structure and function made by us and others, many aspects of its biology are highly unexplored. We will rationally design chemical tools that modulate the activity of ASCT2 using unique mechanisms, including: (A) allosteric inhibitors interacting with a recently identified allosteric site; (B) covalent inhibitors targeting a unique cysteine residue in the substrate binding site of ASCT2; and (C) conformation-specific small molecule modulators targeting specific subpockets in the substrate binding site. In Aim 2, we will characterize SLC6A14, an understudied transporter involved in cancer and metabolic diseases. We will develop structural models of SLC6A14 in different conformations. We will describe biophysical features of the models’ substrate binding site, including electrostatic potential, size, shape, and hydrophobicity, to develop hypotheses for the substrate and inhibitor specificity determinants in SLC6A14. We will use this knowledge to guide the development of inhibitors and substrates, including photoactivatable compounds, to directly test inhibitor-binding site interaction(s). Successful completion of this project will provide a greater understanding of mechanisms of transport and inhi- bition of nutrient transporters, as well as novel chemical tools to further characterize their role in disease. Notably, we will test an emerging and innovative approach to transporter drug discovery that targets allosteric modulation and covalent inhibition via small molecules, to deprive hyper-proliferating cells of nutrients, potentially expanding future applications to treat other diseases that involve SLCs.

总结细胞代谢的改变支持病理学中细胞的快速生长和增殖，例如癌症、Au- 免疫性疾病和心脏缺血，导致对氨基酸代谢的依赖增加，如谷氨酰胺和亮氨酸。营养溶质载体（SLC）转运蛋白在重编程的Meta- 通过为细胞提供用于构建生物量的营养物，bolic网络充当信号分子，增强细胞增殖和分化或调节细胞死亡。我们的主要目标是描述子- 底物和抑制剂特异性决定因素在疾病相关营养SLC转运和发展调节其功能的独特策略。我们采取综合方法，包括计算化学方法，再加上生物化学和生物物理方法和疾病相关的细胞系，以char. aclanine是两种氨基酸转运蛋白，在快速生长细胞的代谢中起关键作用：丙氨酸- 丝氨酸-半胱氨酸转运蛋白（SLC 1A 5，ASCT 2）是一种Na+依赖性氨基酸交换蛋白，可调节细胞内细胞谷氨酰胺水平和氨基酸转运蛋白B 0+（ATB 0+，SLC 6A 14），一种中性和阳离子氨基酸转运蛋白，由Na+和Cl-共同转运驱动。在本项目的目标1中，我们将继续表征ASCT 2，这是一种经过充分验证的用于各种病理学的药物靶点。 GIES（如三阴性乳腺癌和前列腺癌）。尽管最近我们的理解有了进步 ASCT 2的结构和功能，其生物学的许多方面是高度未探索的。我们将合理设计化学工具，使用独特的机制调节ASCT 2的活性，包括： (A)与最近鉴定的变构位点相互作用的变构抑制剂;（B）靶向最近鉴定的变构位点的共价抑制剂; ASCT 2的底物结合位点中的独特半胱氨酸残基;和（C）构象特异性小分子靶向底物结合位点中的特定亚口袋的调节剂。在目标2中，我们将表征SLC 6A 14，一种未被充分研究的与癌症和代谢疾病有关的转运蛋白。我们将开发结构模型不同构象的SLC 6A 14。我们将描述模型底物结合位点的生物物理特征，包括静电势、大小、形状和疏水性，以发展对基底的假设， SLC 6A 14中的抑制剂特异性决定簇。我们将利用这些知识来指导抑制剂的开发和底物，包括可光活化的化合物，以直接测试底物-结合位点相互作用。该项目的成功完成将使人们更好地了解运输和印- 营养转运蛋白的结合，以及新的化学工具，以进一步表征其在疾病中的作用。值得注意的是，我们将测试一种新兴的、创新的方法来发现靶向变构调节的转运药物，和共价抑制通过小分子，剥夺过度增殖细胞的营养，未来应用于治疗涉及SLC的其他疾病。