How do cytosolic proteins interact with lipid droplets? A bio-physical chemical approach to an unresolved biological problem.

肯特州立大学化学系的生命过程化学项目资助了肯特州立大学的埃德加·E·柯伊曼博士和伊丽莎白·K·曼博士，以揭示蛋白质如何与脂滴表面相互作用。脂滴是一种脂肪颗粒，对细胞内能量的供应和调节至关重要，并参与了一系列其他细胞过程。这项研究的重点是细胞中的蛋白质如何靶向并结合到这些脂滴的表面，在那里它们调节液滴的结构和功能。虽然人们对蛋白质如何与包裹复杂细胞的脂膜结合有很多了解，但基本上还没有人知道脂滴是什么。利用模型蛋白，这项研究正在确定与脂滴蛋白结合有关的相互作用。因此，这项工作正在改变人们对脂滴结构和功能的理解。研究生、本科生和高中生在一个高度跨学科的环境中共同工作。此外，这项工作为社会经济背景较低的学生和少数族裔背景的学生带来了有意义的教育经验，目标是在不同的学生中增加STEM学位。本研究项目是关于细胞质脂滴结合蛋白如何与这些关键的、但知之甚少的细胞器表面相互作用的调查。对准确的蛋白质-脂肪相互作用的阐明正在改变对脂滴生物学的理解，因为结果不仅提供了对脂类代谢的洞察，而且还提供了对脂滴的各种功能的洞察。Perilipins，特别是Perilipin 3，作为模型蛋白(S)，准确地描述了不同的蛋白质结构域如何参与脂滴界面的蛋白质-脂肪相互作用。人们特别关注在许多蛋白质中发现的与中性脂质颗粒相互作用的两亲性α-螺旋束结构域。目前正在使用一系列不同的实验工具，包括PI构建的单层槽、PI构建的油滴张力计、定点突变以及蛋白质表达和纯化，并辅之以体内定位研究。因此，这项工作提供了对脂滴结构和生理功能的关键见解。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The Chemistry of Life Processes Program in the Chemistry Division has funded Dr. Edgar E. Kooijman and Dr. Elizabeth K. Mann, at Kent State University, to unravel how proteins interact with the surface of lipid droplets. Lipid droplets are fat particles that are crucial for supplying and regulating energy within the cell, and are involved in a whole host of other cellular processes. This research focuses on how proteins in cells are targeted to and bind to the surface of these lipid droplets, where they regulate droplet structure and function. While much is known about how proteins bind to the lipid membrane that encloses complex cells, essentially nothing is known for lipid droplets. Using model proteins, this research is determining the interactions responsible for the binding of lipid droplet proteins. As such, this work is transforming understanding of the structure and function of lipid droplets. Graduate, undergraduate, and high school students are working together in a highly interdisciplinary environment. Additionally, this work is resulting in meaningful educational experiences for low socioeconomic and minority background students, with the goal of increased STEM degrees among diverse students.This research project is an investigation of how cytosolic lipid droplet binding proteins interact with the surface of these crucial, but poorly understood, organelles. Elucidation of the exact protein-lipid interactions is transforming the understanding of lipid droplet biology as results are providing insight not only on lipid metabolism but also on the diverse functions of lipid droplets. Perilipins, and specifically, perilipin 3, are serving as the model protein(s) to characterize exactly how different protein domains are involved in protein-lipid interaction at the lipid droplet interface. Special attention is being given to the amphipathic alpha-helix bundle domain found in many proteins that interact with neutral lipid particles. A diverse array of experimental tools is being used, including a PI-built monolayer trough, a PI-built oil drop tensiometer, site directed mutagenesis, and protein expression and purification, supplemented by in vivo localization studies. This work thus provides key insights into the structure and physiological function of lipid droplets.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.