PROTEIN PHOSPHORYLATION IN MYCOPLASMA PNEUMONIAE

肺炎支原体中的蛋白质磷酸化

• 批准号: 3148446
• 负责人: DUNCAN C KRAUSE
• 金额: $ 8.14万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 1995-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3148446
• 关键词: Mycoplasma pneumoniae; SDS polyacrylamide gel electrophoresis; antibacterial antibody; autoradiography; bacterial proteins; cell free system; host organism interaction; phosphorus; phosphorylation; protein structure function

Mycoplasma pneumoniae is a member of the class of cell wall-less prokaryotes Mollicutes, which includes the smallest and simplest free- living bacteria known to man. Mycoplasmas have limited biosynthetic capabilities, and many species have resorted to a parasitic lifestyle. As a consequence, mycoplasmas have a great impact on both public health and agriculture. M. pneumoniae is a pathogen of the human respiratory tract and the leading cause of pneumonia in older children and young adults. Adherence by M. pneumoniae to host cells (cytadherence) is pivotal to successful colonization and disease. Mycoplasma protein HMW1 is a phase-variable membrane protein that appears to have an accessory role in cytadherence. HMW1 exhibits an unusual subcellular distribution, being associated specifically with filamentous cell extensions and partitioning primarily, but not exclusively, with the cytoskeleton-like triton shell of M. pneumoniae. This proposal expands upon preliminary data which indicate that HMW1 is phosphorylated. The project addresses fundamental questions regarding the characterization of phosphorylation in vitro and in vivo, as well as the identification and localization of phosphorylated residues in HMW1. Standard techniques will be employed to assess incorporation of 32P into HMW1 during culture and in vitro. HMW1 will be analyzed biochemically to identify phosphorylated residues and localize those residues within the primary amino acid sequence. Finally, the phosphorylation of HMW1 will be examined for a correlation with its partitioning characteristics in Triton X-100 and with mycoplasma cytadherence. The model system offers a number of attractive features, including a target protein associated with an identifiable function (cytadherence), as well as the availability of variants lacking HMW1, of specific antibodies to HMW1, and of deduced amino acid sequence, that is currently being derived from the cloned gene for HMW1. The information gained from these studies will provide the necessary foundation for the long-term objective of defining the significance of HMW1 phosphorylation to protein function.

支原体肺炎是无细胞壁的成员 原核生物软体动物，其​​中包括最小，最简单的自由 - 人类已知的活细菌。 分枝杆菌的生物合成有限 能力和许多物种已诉诸于寄生的生活方式。 结果，支原体对公共卫生都有很大的影响 和农业。 肺炎支原体是人呼吸道的病原体 大小儿童和年轻的肺炎的主要原因和主要原因 成年人。 在 成功定植和疾病关键。 支原体蛋白HMW1 是一种相变的膜蛋白，似乎具有附件 在CytaDherence中的作用。 HMW1表现出异常的亚细胞分布， 特别与丝状细胞扩展和 主要使用细胞骨架样分区（但不是仅限于） 肺炎支原体的Triton壳。 该建议扩展了初步数据，表明HMW1是 磷酸化。 该项目解决了有关的基本问题 体外和体内磷酸化的表征以及 HMW1中磷酸化残基的鉴定和定位。 标准技术将用于评估32p的掺入 HMW1在培养和体外。 HMW1将在生化上进行分析 识别磷酸化的残基并将这些残基定位在 初级氨基酸序列。 最后，HMW1的磷酸化 将检查与其分区特征的相关性 在Triton X-100和支原体中。 模型系统 提供许多吸引人的功能，包括目标蛋白 与可识别函数（CytaDherence）以及 缺乏HMW1的变体的可用性，具有HMW1的特定抗体， 并推导的氨基酸序列，目前是从 HMW1的克隆基因。 从这些研究中获得的信息将 为定义的长期目标提供必要的基础 HMW1磷酸化对蛋白质功能的重要性。