cryoEM Administrative Supplement for Equipment

冷冻电镜设备管理补充文件

• 批准号: 10389992
• 负责人: KENNETH ALLEN TAYLOR
• 金额: $ 24.96万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10389992
• 关键词: Administrative Supplement; Collection; Cryoelectron Microscopy; Data; Defect; Electrons; Equipment; Filament; Funding; Grant; Head; Image; Invertebrates; Length; Mechanics; Muscle; Muscle function; Mutation; Myopathy; Myosin ATPase; Paratropomyosin; Pattern; Proteins; Resolution; Role; Sarcomeres; Striated Muscles; Structure; Tail; Thick Filament; Thin Filament; Three-Dimensional Image; Three-Dimensional Imaging; Ursidae Family; Vertebral column; Vertebrates; connectin; disease-causing mutation; movie; myosin-binding protein C; plasma protein Z; reconstruction; sensor; tool

ABSTRACT Funds are requested for the purchase of a revolutionary new camera, the Apollo, from Direct Electron Ltd. that would enable the collection of large amounts of image data, currently obtained in the form of “movies”. The ability to collect large amounts of data would broaden the types of questions addressable in the PI's R35 grant currently focused on obtaining atomic resolution images of thick filaments from striated muscle. Generally, thick filaments from all striated muscles have four structurally distinct regions: (1) a 160 nm long bare zone in the center where myosin tails are packed antiparallel to each other, (2) an A-band follows where myosin molecules are arranged in a helical or quasi helical structure with the myosin tails packed in a parallel arrangement, (3) a tapered end where the connecting filament, e.g. titin, connects the thick filament to the Z-disk, and (4) features along the thick filament A-band from invertebrates in particular, but also from vertebrates, that fail to follow the myosin molecule arrangement. The helical arrangement of myosin in the A-band of invertebrate thick filaments facilitates an atomic structure of the myosin tail and head arrangement by using all of the myosin axial repeats from the end of the bare zone to the beginning of the tapered ends. However, the bare zone, the transition from bare zone to helical arrangement, the tapered ends as well as additional features that do not follow the myosin helical symmetry perform important roles in the assembly of thick filaments. Features, such as the paramyosin of invertebrate thick filaments fail to follow the myosin helical symmetry in the A-band and are poorly represented in cryoEM reconstructions. Paramyosin in concert with other non-myosin proteins may function to determine the thick filament length in invertebrates. Titin, myosin binding protein C and other non-myosin proteins of vertebrate thick filaments also do not follow a regular pattern along the entire A-band. Acquisition of a much faster camera capable of recording a 60-frame movie in a single second, will provide the tool needed to obtain high resolution 3-D images of these features that are not well represented if at all in structures solved by enforcing helical symmetry.

抽象的 要求从Direct购买革命性的新相机Apollo的资金 Electron Ltd.将使目前获得的大量图像数据收集 以“电影”的形式。收集大量数据的能力将扩大 PI的R35赠款中可解决的问题目前致力于获得原子分辨率 浓密的肌肉厚细丝的图像。通常，所有条纹的厚细丝 肌肉有四个结构上不同的区域：（1）中心的160 nm长裸露区域 肌球蛋白尾巴彼此挤压，（2）A频段跟随肌球蛋白 分子以螺旋或准螺旋结构排列，肌球蛋白尾巴包装在 平行排列，（3）连接丝，例如titin，连接 Z盘的细丝厚，（4）沿无脊椎动物的厚丝a频段的特征 特别是，也从脊椎动物中，无法遵循肌球蛋白分子排列。这 肌球蛋白在无脊椎动物厚细丝的A带中的螺旋排列有助于原子 肌球蛋白尾巴和头部排列的结构，使用所有肌球蛋白轴向重复 裸露区域的末端到锥形末端的开头。但是，裸露区域， 从裸露区域到螺旋排列的过渡，锥形末端以及其他功能 不遵循肌球蛋白螺旋对称性在厚的组装中起重要作用 细丝。特征，例如无脊椎动物厚细丝的副蛋白，无法遵循 A波段中的肌球蛋白螺旋对称性，在冷冻重建中表示不良。 与其他非肌球蛋白蛋白协同合作的帕莫色素可能起作用以确定厚 无脊椎动物的细丝长度。滴定，肌球蛋白结合蛋白C和其他非肌球蛋白蛋白 脊椎动物厚的细丝也不会沿着整个A波段遵循常规图案。 收购能够在一秒钟内录制60帧电影的速度更快的相机 将提供获得这些功能的高分辨率3-D图像所需的工具 如果通过强制执行螺旋对称性解决的结构，则可以很好地表示。