Molecular mechanism of the NKCC transporter

NKCC转运蛋白的分子机制

• 批准号: 9395166
• 负责人: Thomas Chew
• 金额: $ 3.75万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2021-12-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9395166
• 关键词: Address; Adverse effects; Antibodies; Basic Science; Binding; Binding Sites; Biological Assay; Body part; Bumetanide; Cell membrane; Chlorthalidone; Cirrhosis; Clinic; Clinical; Coupled; Coupling; Data; Dehydration; Development; Disease; Diuretics; Drug Interactions; Drug Targeting; Edema; Environment; Event; Family; Family member; Furosemide; Goals; Heart failure; Homeostasis; Hydrochlorothiazide; Hypertension; Hypokalemia; In Vitro; Individual; Interdisciplinary Study; Ion Transport; Ions; Kidney; Knowledge; Lipid Bilayers; Lipids; Liposomes; Medicine; Membrane Proteins; Mentors; Modification; Molecular; Molecular Conformation; Mutagenesis; Negative Staining; Nephrotic Syndrome; Particle Size; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phosphorylation; Physiology; Post-Translational Protein Processing; Potassium Chloride; Property; Protein Family; Proteins; Research; Resolution; Role; SLC12A3 gene; Sampling; Sodium; Sodium Chloride; Sodium-Potassium-Chloride Symporters; Specificity; Structure; Substrate Specificity; System; Techniques; Thiazide Diuretics; blood pressure regulation; computer studies; design; drug development; experience; experimental study; extracellular; glycosylation; graduate student; hearing impairment; inhibitor/antagonist; insight; member; particle; protein function; protein structure; reconstitution; solute; stoichiometry

Abstract The sodium-potassium-chloride-cotransporter (NKCC) family of transporters are critical to the reabsorption of ions in the kidney and other parts of the body. As a member of the larger solute carrier 12 (SLC12A) family, these transporters move sodium, potassium, and chloride across the cell membrane in order to maintain ionic homeostasis and regulate blood pressure. As a result of their important roles in physiology, they are also among the most widely targeted transporters in medicine. Drugs such as the loop diuretics and thiazide diuretics have been in use for decades, and are fundamental medications to treating conditions such as edema, cirrhosis, heart failure, and hypertension. Yet despite their widespread use, little is known about the molecular mechanism by which they operate. This project will address that gap in knowledge, by studying the NKCC group of transporters from a fundamental, basic science perspective. An in vitro functional assay will be developed in order to study the transport mechanism of NKCC in a well-defined system. Key components responsible for ion recognition and binding, as well as inhibitor binding sites will be determined using mutagenesis coupled with functional assays. The above knowledge will be combined with structural characterizations about NKCC to understand its function. As a whole, this study will reveal the fundamental mechanisms about how NKCC transporters operate and how they are targeted. This information will lead to better pharmaceuticals, with fewer side effects.

摘要 钠-钾-氯化物-协同转运蛋白（NKCC）家族转运蛋白对药物的重吸收至关重要。 肾脏和身体其他部位的离子。作为较大溶质载体12（SLC 12 A）家族的成员， 这些转运蛋白将钠、钾和氯化物穿过细胞膜以维持离子状态 体内平衡和调节血压。由于它们在生理学中的重要作用，它们也 是医学上最广泛的靶向转运蛋白之一。袢利尿剂和噻嗪类药物 利尿剂已经使用了几十年，并且是治疗以下病症的基本药物， 水肿、肝硬化、心力衰竭和高血压。然而，尽管它们被广泛使用，但人们对它们知之甚少。 它们运作的分子机制。本项目将通过研究 NKCC运输集团从一个基本的，基础科学的角度。将进行体外功能测定， 为了研究NKCC在一个定义明确的系统中的传输机制而开发的。关键部件 负责离子识别和结合以及抑制剂结合位点将使用 诱变结合功能测定。上述知识将结合结构 对NKCC的描述，以了解其功能。作为一个整体，这项研究将揭示基本的 关于NKCC运输者如何运作以及他们如何成为目标的机制。这些信息将导致 更好的药物，副作用更少。