Equipment Supplement R01GM126626: Mechanistic studies of prokaryotic and eukaryotic nitrate/nitrite transport

设备补充 R01GM126626：原核和真核硝酸盐/亚硝酸盐运输的机制研究

• 批准号: 10377771
• 负责人: Hongjin Zheng
• 金额: $ 7.19万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377771
• 关键词: Blood Circulation; Cancerous; Cellular Membrane; Communities; Crystallization; Equipment; Escherichia coli; Event; Genetic Diseases; Health; Homeostasis; Human; Knowledge; Life; Light; Mediating; Membrane Proteins; Molecular; Molecular Conformation; Mutagenesis; Nitrates; Nitric Oxide; Nitrites; Nitrogen; Oral cavity; Pharmacologic Substance; Play; Process; Resolution; Role; Salivary Glands; Structure; Structure-Activity Relationship; Time; commensal bacteria; drug development; flexibility; nitrate transporter; uptake

Recent studies suggest that nitrate and nitrite molecules have profound beneficial effects to human health, though they were thought to be cancerous since 1970s. To maximize their pharmaceutical potential, we need to first understand how nitrate and nitrite circulate in humans. Such circulation depends on two critical events: active accumulation of nitrate mediated by sialin transporters in salivary glands, as well as nitrate uptake and nitrite secretion by commensal bacteria in the mouth. Thus, nitrate/nitrite molecules have to cross different cellular membranes multiple times, before being used by humans. These translocation processes are mediated by a group of membrane proteins called nitrate transporters. To understand how these transporters function, we solved high-resolution crystal structures of NarK from E. coli, and further demonstrated that, surprisingly, NarK is a nitrate/nitrite exchanger. Despite the progress we made, the detailed molecular mechanisms of nitrate/nitrite translocation are still largely unknown. In this proposal, we aim to fill the knowledge gap by: 1) understand substrate selectivity and conformational flexibility using directed-mutagenesis of NarK, so to better understand the function of NarK at the molecular level; 2) obtain high-resolution structures of NarK in previously unobserved conformation, so we can reconstruct the complete transport cycle of NarK; 3) explore the structure-function relationship of human nitrate transporter sialin, so we will understand the similarities and differences among nitrate transporters from diverse species. Overall, upon completion of the proposal, we expect to expand our general understanding of the nitrate/nitrite transport and shed light on the crucial roles that nitrate transporters (both eukaryotic and prokaryotic) play in nitrate/nitrite circulation. The knowledge gained here will facilitate potential drug development related to bacterial nitrate transporters and human sialin.

最近的研究表明，硝酸盐和亚硝酸盐分子对人类有深远的有益影响 健康，尽管自20世纪70年代以来，它们被认为是癌症。最大限度地利用他们的药品 潜在的，我们需要首先了解硝酸盐和亚硝酸盐如何在人体内循环。这样的流通 取决于两个关键事件：唾液淋巴转运蛋白介导的硝酸盐在唾液中的活跃积累 口腔中共生细菌对硝酸盐的摄取和亚硝酸盐的分泌。因此， 硝酸盐/亚硝酸盐分子在使用之前必须多次穿过不同的细胞膜 被人类发现。这些转位过程是由一组膜蛋白介导的，称为 硝酸盐转运体。为了了解这些转运蛋白是如何工作的，我们解决了高分辨率晶体 的结构，进一步证明了，令人惊讶的是，它是一种硝酸盐/亚硝酸盐。 交易所。尽管我们取得了进展，但硝酸盐/亚硝酸盐的详细分子机制 易位在很大程度上仍然是未知的。在本提案中，我们的目标是通过以下方式填补知识缺口：1) 通过NAK的定向突变了解底物的选择性和构象灵活性，因此 在分子水平上更好地了解nark的功能；2)获得高分辨结构 以前没有观察到的构象，所以我们可以重建完整的运输循环 3)探索人类硝酸盐转运蛋白sialin的结构-功能关系，因此我们将 了解不同物种硝酸盐转运蛋白的异同。总的来说， 建议完成后，我们期望扩大我们对硝酸盐/亚硝酸盐的一般了解。 运输并阐明硝酸盐转运蛋白(真核和原核生物)的关键作用 在硝酸盐/亚硝酸盐循环中发挥作用。在这里获得的知识将促进潜在的药物开发 与细菌硝酸盐转运蛋白和人类唾液酸有关。

项目成果

Mechanosensitive channel YnaI has lipid-bound extended sensor paddles.

机械敏感通道 YnaI 具有脂质结合的扩展传感器板。

• DOI: 10.1038/s42003-021-02122-0
• 发表时间: 2021-05-20
• 期刊: Communications biology
• 影响因子: 5.9
• 作者: Hu W;Wang Z;Zheng H
• 通讯作者: Zheng H

Pathogenic siderophore ABC importer YbtPQ adopts a surprising fold of exporter.

致病性铁载体 ABC 输入蛋白 YbtPQ 采用了令人惊讶的输出蛋白折叠。

• DOI: 10.1126/sciadv.aay7997
• 发表时间: 2020
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Wang,Zhiming;Hu,Wenxin;Zheng,Hongjin
• 通讯作者: Zheng,Hongjin

The molecular mechanism of sialic acid transport mediated by Sialin.

• DOI: 10.1126/sciadv.ade8346
• 发表时间: 2023-01-20
• 期刊: SCIENCE ADVANCES
• 影响因子: 13.6
• 作者: Hu, Wenxin;Chi, Congwu;Song, Kunhua;Zheng, Hongjin
• 通讯作者: Zheng, Hongjin

Structural and functional diversity calls for a new classification of ABC transporters.

• DOI: 10.1002/1873-3468.13935
• 发表时间: 2020-12
• 期刊: FEBS letters
• 影响因子: 3.5
• 作者: Thomas C;Aller SG;Beis K;Carpenter EP;Chang G;Chen L;Dassa E;Dean M;Duong Van Hoa F;Ekiert D;Ford R;Gaudet R;Gong X;Holland IB;Huang Y;Kahne DK;Kato H;Koronakis V;Koth CM;Lee Y;Lewinson O;Lill R;Martinoia E;Murakami S;Pinkett HW;Poolman B;Rosenbaum D;Sarkadi B;Schmitt L;Schneider E;Shi Y;Shyng SL;Slotboom DJ;Tajkhorshid E;Tieleman DP;Ueda K;Váradi A;Wen PC;Yan N;Zhang P;Zheng H;Zimmer J;Tampé R
• 通讯作者: Tampé R

Mechanism and inhibition of Streptococcus pneumoniae IgA1 protease.

• DOI: 10.1038/s41467-020-19887-3
• 发表时间: 2020-11-27
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Wang Z;Rahkola J;Redzic JS;Chi YC;Tran N;Holyoak T;Zheng H;Janoff E;Eisenmesser E
• 通讯作者: Eisenmesser E