NONRANDOM UNFOLDED STATES AND PROTEIN STABILITY

非随机展开状态和蛋白质稳定性

• 批准号: 2713749
• 负责人: TIMOTHY M LOGAN
• 金额: $ 10.79万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2713749
• 关键词: conformation; peptidylprolyl isomerase; protein folding; spectrometry; thermodynamics

DESCRIPTION: The objective of this research program is to seek a greater understanding of the significance of non-random structure present in the unfolded states of proteins for protein stability and folding. The FK506 binding protein (FKBP) provides a unique system to identify such correlations. High resolution structures of FKBP in both the crystal and solution forms are available and the equilibrium unfolding of FKBP has been thoroughly characterized using a variety of spectroscopic and biochemical methods. In addition, a detailed structural study of FKBP unfolded in concentrated urea and guanidine hydrochloride has been performed, demonstrating the presence of non-random structure associated with specific residues. The location of the non-random structure in unfolded FKBP was correlated with similar structures observed in the folded form, and was more strongly correlated with the propensity for helical and turn conformations predicted from statistical and thermodynamic scales of secondary structure prediction. On the other hand, a particularly surprising finding from these initial studies was that different secondary structures are formed in the folded and unfolded states for the C-terminal residues. Mutagenesis and spectroscopic approaches will be combined to further characterize the unfolded form of FKBP and to explore these relationships in detail. The previous structural and thermodynamic studies of folded and unfolded FKBP will be used to understand the role that non-random structure in the unfolded state plays in the folding and stability of FKBP. The role of non-random structures in altering native state stability in FKBP will identify general rules for similar structures in other proteins. In addition, deciphering the thermodynamic driving forces responsible for the conformational switch in the C-terminal residue upon folding will have implications in protein design and in three dimensional structure prediction.

描述：这项研究计划的目标是寻求一个更大的 理解非随机结构的重要性， 蛋白质的非折叠状态，以实现蛋白质的稳定性和折叠。 的fk 506 结合蛋白（FKBP）提供了一个独特的系统，以确定这种 相关性 FKBP在晶体和晶体中的高分辨率结构 解的形式是可用的，FKBP的平衡展开具有 使用各种光谱和 生物化学方法 此外，FKBP的详细结构研究 在浓尿素和盐酸胍中展开， 进行，证明存在非随机结构相关 特定的残留物。 非随机结构在 未折叠的FKBP与在细胞中观察到的类似结构相关。 折叠的形式，并与更强烈的倾向， 从统计和 二级结构预测的热力学尺度。 另 一方面，这些初步研究中一个特别令人惊讶的发现是， 折叠和展开后形成不同的二级结构 C-末端残基的状态。 诱变和光谱 将结合各种方法进一步描述展开形式的特征 并详细探讨这些关系。 折叠和未折叠的分子的结构和热力学研究 FKBP将用于理解非随机结构在 未折叠状态对FKBP的折叠和稳定性起作用。 的作用 非随机结构在改变FKBP中的自然状态稳定性方面的作用将 确定其他蛋白质中类似结构的一般规则。 在 此外，破译的热力学驱动力负责 折叠时C-末端残基中的构象转换将 在蛋白质设计和三维结构中有着重要的意义 预测.