Studies on the Protein-assisted Mechanism for Intracellular Membrane Contact Sites

细胞内膜接触位点的蛋白质辅助机制研究

• 批准号: 1951425
• 负责人: Jeffery Klauda
• 金额: $ 98.38万
• 依托单位: University of Maryland, College Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1951425&HistoricalAwards=false
• 关键词: Studies; Protein; assisted; Mechanism; Intracellular

This project will probe the mechanism of lipid movement between cellular compartments known as organelles. Recently, strong evidence indicates that most lipid transport occurs with the help of lipid transport proteins that can facilitate the formation of organelle contacts known as membrane contact sites. The oxysterol binding homologue (Osh) proteins of yeast are an example of lipid transport proteins that form membrane contact sites and serve as an excellent model for lipid exchange between organelles. Since cells can be used to produce chemicals for commercial use, understanding lipid exchange and ways to modify it could improve production of chemicals. This project will provide scientific training for researchers with focused recruitment from underrepresented groups. Project concepts and results will be used in educational outreach for minority-serving high school summer programs, courses, and developing hands-on protein engineering activities for the general public. Osh proteins were originally thought to transport sterols but have recently been shown to be important in the transport of signaling lipids, such as phosphatidylinositols. This project integrates experimental and computational techniques to elucidate membrane contact site formation and probe lipid exchange by first studying membrane-binding regions of Osh proteins and then the mechanism the full-length protein uses to form membrane contact sites. The binding of the lipid-packing sensor of Osh proteins to model membranes (planar and curved) will be probed using enhanced sampling simulations, neutron diffraction, and high-resolution field-cycling NMR. Other domains of Osh4 interacting with membranes will be studied with a focus on how this protein might conformationally change to facilitate membrane contact sites. Ultimately, the mechanism of the full-length Osh4 protein to form membrane contact sites and lipid transfer will be probed by developing new computational tools for dual-membrane binding and the use of field-cycling NMR and fluorescence techniques. This proposal is jointly funded by the Cellular Dynamics and Function cluster and Molecular Biophysics cluster in the Division of Molecular and Cellular Biosciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该项目将探讨脂质在细胞器之间的运动机制。最近，强有力的证据表明，大多数脂质转运是在脂质转运蛋白的帮助下发生的，脂质转运蛋白可以促进细胞器接触的形成，即膜接触位点。酵母的氧甾醇结合同源（Osh）蛋白是脂质转运蛋白的一个例子，它可以形成膜接触位点，并作为细胞器之间脂质交换的一个很好的模型。由于细胞可以用于生产商业用途的化学品，了解脂质交换和修改它的方法可以改善化学品的生产。该项目将为研究人员提供科学培训，重点从代表性不足的群体中招募人员。项目概念和成果将用于为少数族裔服务的高中暑期项目和课程的教育推广，并为公众开发动手蛋白质工程活动。奥什蛋白最初被认为是运输甾醇，但最近被证明在运输信号脂质（如磷脂酰肌醇）中起重要作用。本项目结合实验和计算技术，首先研究Osh蛋白的膜结合区，然后研究全长蛋白形成膜接触位点的机制，阐明膜接触位点的形成和脂质交换的探测。Osh蛋白的脂质填充传感器与模型膜（平面和弯曲）的结合将使用增强的采样模拟、中子衍射和高分辨率场循环核磁共振进行探测。Osh4与膜相互作用的其他结构域将被研究，重点是该蛋白如何改变构象以促进膜接触位点。最终，我们将通过开发新的双膜结合计算工具以及使用场循环核磁共振和荧光技术来探索全长Osh4蛋白形成膜接触位点和脂质转移的机制。本提案由分子和细胞生物科学部的细胞动力学和功能集群和分子生物物理集群共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Modeling the membrane binding mechanism of a lipid transport protein Osh4 to single membranes

模拟脂质转运蛋白 Osh4 与单膜的膜结合机制

• DOI: 10.1016/j.bpj.2022.03.001
• 发表时间: 2022
• 期刊: Biophysical Journal
• 影响因子: 3.4
• 作者: Karmakar, Sharmistha;Klauda, Jeffery B.
• 通讯作者: Klauda, Jeffery B.

Considerations of Recent All-Atom Lipid Force Field Development

• DOI: 10.1021/acs.jpcb.1c02417
• 发表时间: 2021-05-28
• 期刊: JOURNAL OF PHYSICAL CHEMISTRY B
• 影响因子: 3.3
• 作者: Klauda, Jeffery B.

Estimating localization of various statins within a POPC bilayer

• DOI: 10.1016/j.chemphyslip.2021.105074
• 发表时间: 2021-03-10
• 期刊: CHEMISTRY AND PHYSICS OF LIPIDS
• 影响因子: 3.4
• 作者: Kuba, Jacob Olondo;Yu, Yalun;Klauda, Jeffery B.
• 通讯作者: Klauda, Jeffery B.

All-Atom Modeling of Complex Cellular Membranes

• DOI: 10.1021/acs.langmuir.1c02084
• 发表时间: 2021-12-28
• 期刊: LANGMUIR
• 影响因子: 3.9
• 作者: Hsieh, Min-Kang;Yu, Yalun;Klauda, Jeffery B.
• 通讯作者: Klauda, Jeffery B.

Molecular dynamics simulations of the human ocular lens with age and cataract

• DOI: 10.1016/j.bbamem.2022.184025
• 发表时间: 2022-08-17
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
• 影响因子: 3.4
• 作者: Fernandes, Joshua B.;Yu, Yalun;Klauda, Jeffery B.
• 通讯作者: Klauda, Jeffery B.