EAPSI: Understanding the Contribution of Class I Myosins to Cellular Processes Involving Membrane Deformation

EAPSI：了解 I 类肌球蛋白对涉及膜变形的细胞过程的贡献

• 批准号: 1515223
• 负责人: Sarah Barger
• 金额: $ 0.51万
• 依托单位: Barger Sarah R
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1515223&HistoricalAwards=false
• 关键词: EAPSI; Understanding; Contribution; Class; Myosins

Plant cells are bound by a rigid cell wall, yet animal cells rely only on a plasmamembrane and the underlying meshwork of cytoskeletal fibers, called actin filaments, for support and stability. As the cell boundary, the plasma membrane both enables and restricts any shape changing process. Adherence of the plasma membrane to the actin meshwork beneath generates resistance to deformation but also allows cells to maintain complex shapes. Non-muscle Class I myosins are motor proteins capable of binding both plasma membrane and actin filaments; therefore, they may link the plasma membrane to the actin cytoskeleton. That Class I myosins are structurally conserved from yeast to humans is indicative of their functional importance. However, their precise role in processes involving membrane deformation, such as cell migration and phagocytosis, remains elusive. In collaboration with Dr. Nils Gauthier, an expert in membrane-cytoskeletal interactions at the Mechanobiology Institute at the National University of Singapore, this research will evaluate the participation of Class I myosins in membrane-cytoskeletal adhesion during physiologically important processes that involve membrane deformation. In this project, the strength of membrane-cytoskeletal attachment in macrophages (cells of the innate immune system) lacking Class I myosins will be measured using a calibrated optical tweezers system, which uses a focused laser to measure forces applied to microscopic objects. Cells will then be subjected to tests on cell motility, cell traction and cell spreading to assess membrane deformation ability. These experiments will also be performed on macrophages expressing truncated myosin constructs to further our understanding of the roles of the individual protein domains. With the expertise of Dr. Gauthier and the specialized core facilities of the Mechanobiology Institute, this work will shed light on the workings of an ancient motor protein and its role in moving the plasma membrane. This award is funded in collaboration with the National Research Foundation of Singapore.

植物细胞由刚性细胞壁结合，但动物细胞仅依赖于浆膜和细胞骨架纤维（称为肌动蛋白丝）的潜在网状作品，以进行辅助和稳定性。 作为细胞边界，质膜既可以启用并限制任何形状变化的过程。 质膜粘附在下面的肌动蛋白网晶中产生抗变形性，但也使细胞保持复杂的形状。 非肌肉I类是运动蛋白，能够结合质膜和肌动蛋白丝。因此，它们可能将质膜连接到肌动蛋白细胞骨架。 从酵母到人类，I级肌动物在结构上是保守的，这表明它们的功能重要性。 但是，它们在涉及膜变形的过程中的精确作用，例如细胞迁移和吞噬作用，仍然难以捉摸。 与新加坡国立大学机械生物学研究所的膜 - 细胞骨架相互作用的专家Nils Gauthier博士合作，该研究将评估I类肌球蛋白参与涉及膜变形的膜上重要过程中膜 - 细胞骨架骨骼粘附的参与。 在这个项目中，将使用校准的光学镊子系统测量缺乏I类肌球蛋白的巨噬细胞（先天免疫系统）中膜细胞骨架附着的强度，该系统使用聚焦激光器来测量应用于微观物体的力。 然后，细胞将经过细胞运动，细胞牵引和细胞扩散的测试，以评估膜变形能力。 这些实验还将对表达截短的肌球蛋白构建体的巨噬细胞进行，以进一步了解我们对单个蛋白质结构域的作用的理解。 借助Gauthier博士的专业知识和机械生物学研究所的专业核心设施，这项工作将阐明古老的运动蛋白的运作及其在移动质膜中的作用。该奖项由与新加坡国家研究基金会合作资助。