Membrane Protein Stability

膜蛋白稳定性

• 批准号: 7103700
• 负责人: JAMES U BOWIE
• 金额: $ 27.26万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7103700
• 关键词: Escherichia coli; Halobacteriaceae; X ray crystallography; bacterial genetics; bacteriorhodopsins; calorimetry; chemical kinetics; chemical stability; crystallization; enzyme induction /repression; gene mutation; hydrogen bond; mass spectrometry; membrane proteins; model design /development; permease; phosphotransferases; physical model; point mutation; protein folding; protein structure function; racemization; solubility; structural biology; thermodynamics

DESCRIPTION (provided by applicant): Knowledge of energetic contributions to protein folding is essential for many of the central quests of structural biology, including protein design and modeling, as well as an understanding of protein function and dysfunction in disease. Consequently, an enormous effort has been made to work out the forces that stabilize protein structure. In this effort, membrane proteins have been largely neglected, in spite of the fact that they play critical biological roles and are the major targets of drugs. Membrane protein folding has been avoided because of the enormous complexity of the membrane environment and the technical challenges faced when studying this problem. In the prior grant period, we have overcome many of these challenges and are finally well positioned to dissect the energetics of folding in a large membrane protein. To my knowledge, there is no other group that combines both thermodynamic stability measurements and detailed structural analysis in a similar way. The Specific Aims are: I.To probe packing forces, we will introduce bumps and create holes at various points in the bacteriorhodopsin (bR) structure and examine the consequences for bR structure and stability. II. To probe hydrogen bond strength, we will specifically delete inter-helical hydrogen bonds in bR and examine the consequences for bR structure and stability. III. To probe the contribution of interhelical loops, we will examine the consequences of side chain deletions and loop insertions on bR structure and stability. IV. Develop hydrogen exchange as a means to probe the unfolded state of bR.

描述（由申请人提供）：对蛋白质折叠的能量贡献的知识对于许多结构生物学的主要追求，包括蛋白质设计和建模以及对蛋白质功能和疾病功能障碍的理解至关重要。因此，已经付出了巨大的努力来解决稳定蛋白质结构的力。在这项工作中，尽管它们起着关键的生物学作用，并且是药物的主要靶标，但膜蛋白已被大大忽略了。由于膜环境的巨大复杂性以及研究此问题时面临的技术挑战，因此避免了膜蛋白折叠。在以前的赠款期间，我们已经克服了许多挑战，并最终在大型膜蛋白中的折叠能量学方面做到了很好的位置。据我所知，没有其他群体以类似的方式结合了热力学稳定性测量和详细的结构分析。 具体目的是： I.在探测堆积力的情况下，我们将在细菌紫红素（BR）结构中引入凸起并在各个点创建孔，并检查BR结构和稳定性的后果。 ii。为了探测氢键强度，我们将特异性删除BR中的螺旋间氢键，并检查BR结构和稳定性的后果。 iii。为了探测螺旋间环的贡献，我们将研究侧链缺失和循环插入对BR结构和稳定性的后果。 iv。开发氢交换，以此作为探测BR的展开状态的一种手段。