HEAT SHOCK PROTEIN HSP-90 FOLLOWING INJURY

受伤后的热休克蛋白 HSP-90

• 批准号: 2027119
• 负责人: MICHAEL B YAFFE
• 金额: $ 7.8万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2027119
• 关键词: biological signal transduction; chemical binding; enzyme structure; histopathology; intermolecular interaction; molecular chaperones; molecular pathology; protein kinase; protein structure function; stress proteins; tissue /cell culture

DESCRIPTION (Adapted from the applicant's abstract) The regulation of signal transduction events following tissue injury is of prime importance in maintaining cell viability. This process is particularly relevant to diseases of the respiratory and cardiovascular system, where tissue insults such as ischemia, reperfusion, and sepsis are common. The applicants propose that this occurs through the involvement of the 90-kDa heat-shock protein hsp9O. Hsp9O is an abundant and ubiquitously distributed stress protein present in all living organisms from bacteria to man. Although hsp9O plays a role in steroid receptor maturation, and forms stable associations with several protein kinases, no clear picture regarding hsp9O's exact function at the molecular level has emerged. In this proposal, they hypothesize that hsp9O functions as a protein kinase chaperone, assisting in the correct packing of the regulatory and catalytic domains. By sequestering the catalytic subunits of a variety of protein kinases following injury, hsp9O could prevent both non-specific kinase denaturation/aggregation and the inappropriate activation of signal transduction pathways. In vivo cell culture experiments will be used to determine the identity and activity of protein kinases that interact with hsp9O following injury. In vitro protein biochemistry experiments will examine which kinase subfamilies contain members capable of interacting with hsp9O, and elucidate whether hsp9O plays a chaperone-like function in this interaction. Finally the mechanism of hsp9O kinase binding and release will be investigated using kinase mutants, domain- and loop-swapping constructs, and oriented peptide libraries, based on extrapolations from known protein kinase crystal structures.

描述 （改编自申请人摘要）信号的调节 组织损伤后的转导事件对于 维持细胞活力。 这一进程特别关系到 呼吸和心血管系统疾病，其中组织损伤 如局部缺血、再灌注和脓毒症是常见。 申请人 我认为这是通过参与90 kDa的热休克发生的， hsp 90蛋白。 Hsp 9 O是一种丰富的、广泛分布的应激蛋白， 蛋白质存在于从细菌到人类的所有生物体中。虽然 hsp 90在类固醇受体成熟中起作用，并形成稳定的 与几种蛋白激酶的关联，没有关于 hsp 9 O在分子水平上的确切功能已经显现。 在这 他们假设hsp 9 O作为一种蛋白激酶发挥功能， 伴侣，协助正确包装的监管和催化 域. 通过隔离多种蛋白质的催化亚基 损伤后，hsp 9 O可以阻止两种非特异性激酶 变性/聚集和信号的不适当激活 转导途径。 体内细胞培养实验将用于确定同一性和 损伤后与hsp 90相互作用的蛋白激酶的活性。 在 体外蛋白质生物化学实验将检查哪些激酶亚家族 包含能够与hsp 9 O相互作用的成员，并阐明是否 hsp 90在这种相互作用中起着类似伴侣的作用。 最后 hsp 9 O激酶结合和释放的机制将使用 激酶突变体、结构域和环交换构建体和定向肽 基于已知蛋白激酶晶体外推的文库 结构.