Microsequencing of a Putatve Myosin I Isoform

Putatve 肌球蛋白 I 亚型的微测序

• 批准号: 9631833
• 负责人: Primal de Lanerolle
• 金额: $ 5万
• 依托单位: University of Illinois at Chicago
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 1998-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9631833&HistoricalAwards=false
• 关键词: Microsequencing; Putatve; Myosin; Isoform

9631833 de Lanerolle The term "myosin" describes a family of motor proteins that are characterized by their ability to hydrolyze ATP when they bind actin. Myosin II, the most familiar and best characterized member of this family, is a filamentous protein that uses the energy released when ATP is hydrolyzed to move filaments past each other, an essential step in muscle contraction and in cytokinesis. In contrast to our understanding of the physiological properties of myosin II, little is known about the other members of this superfamily, many of which do not form filaments. Of these non-filamentous myosins, myosin I has been studied most extensively. Myosin I was initially described in Acanthamoeba and is now known to be widely distributed in mammalian cells. This protein has been studied extensively at the molecular and biochemical levels, and there is an abundance of information about the cellular distribution of myosin I. The physiological role of this protein, particularly in mammalian cells, is less well defined. In an effort the investigate the physiological role of myosin I in mammalian cells, the protein was purified from bovine adrenal glands and a polyclonal antibody was raised against it. While characterizing these antibodies, it was observed that the affinity-purified antibodies to myosin I stained the nucleus in interphase cells. In light of the recent demonstration by others of actin-like proteins in the nucleus, this data prompted the hypothesis that myosin I is involved in nuclear functions. However, before such a putative role can be investigated, the protein in the nucleus which reacts with the antibody must be unambiguously identified. Therefore, the goal of this research is to purify sufficient protein to identify it by microsequencing it. ***

小行星9631833 术语“肌球蛋白”描述了一个家族的马达蛋白，其特征在于当它们结合肌动蛋白时水解ATP的能力。 肌球蛋白II是这个家族中最熟悉和最具特征的成员，是一种丝状蛋白质，它利用ATP水解时释放的能量使细丝相互移动，这是肌肉收缩和胞质分裂的重要步骤。 与我们对肌球蛋白II生理特性的理解相反，我们对这个超家族的其他成员知之甚少，其中许多成员不形成肌丝。 在这些非丝状肌球蛋白中，肌球蛋白I已被最广泛地研究。 肌球蛋白I最初在阿米巴中被描述，现在已知广泛分布于哺乳动物细胞中。 这种蛋白质已经在分子和生物化学水平上进行了广泛的研究，并且有大量关于肌球蛋白I细胞分布的信息。 这种蛋白质的生理作用，特别是在哺乳动物细胞中，是不太清楚。 为了研究肌球蛋白I在哺乳动物细胞中的生理作用，从牛肾上腺中纯化了肌球蛋白I，并制备了抗肌球蛋白I的多克隆抗体，在鉴定这些抗体时，观察到亲和纯化的肌球蛋白I抗体染色间期细胞的细胞核。 根据最近其他人对细胞核中肌动蛋白样蛋白的证明，这些数据促使人们提出肌球蛋白I参与核功能的假设。 然而，在研究这种假定的作用之前，必须明确鉴定细胞核中与抗体反应的蛋白质。 因此，本研究的目标是纯化足够的蛋白质，通过微测序对其进行鉴定。