Development and Application of Methods for Investigating the Biological Roles and Structural Basis of Protein Kinetic Stability

点击翻译按钮获取中文摘要

Intellectual MeritLike a child strapped to a high-chair, some proteins are trapped in their native structure, unable to access partially and globally unfolded conformations. This may be a method used by "mother nature" to protect certain proteins from premature degradation, which would compromise the well being of the organism. Proteins that possess this property are described as being kinetically stable because, unlike most proteins, their long "shelf-life" and stability towards degradation is kinetically controlled by its very slow unfolding rate. The structural basis and biological significance of protein kinetic stability remains poorly understood. Therefore, the main goal of this project is to gain novel insight into the role of kinetically stable proteins (KSPs) in the survival and adaptation of prokaryotic thermophiles and mesophiles, and to gain enhanced understanding about the structural basis of kinetic stability. The central dogma is that bacteria possess KSPs with functions that are essential to survive in their respective environments. Novel electrophoresis methods developed in the laboratory of the Principle Investigator will be further developed and applied to: 1) characterize the proteome of KSPs in several thermophilic and mesophilic bacteria; 2) quantify the kinetic stability of hyperstable proteins directly in the cellular lysate of bacteria; and 3) construct an accessible online database of KSPs to explore the structural and functional basis of kinetic stability. It is expected that the results of this project will enhance the understanding of the biological significance of kinetic stability, and will provide a convenient and accessible assay to quantify the kinetic stability of proteins in their biological milieu. Furthermore, it will yield a wealth of accessible data that will stimulate studies to understand the biological roles of KSPs in the survival, adaptation, and evolution of thermophilic and mesophilic organisms. Broader ImpactAs a broader impact, this project will lead to the discovery of KSPs with potential for biotechnology applications of benefit to society. In terms of education, this project will involve postdoctoral, graduate, and undergraduate researchers working together in a stimulating interdisciplinary environment that integrates training and learning. The Principal Investigator is developing a local program (RISE: Research Internship for Community College Student Engagement) for local community college students that show interest in STEM disciplines and have the academic potential and desire to transfer to a four-year university. A RISE student will participate every summer in the project and will receive mentoring and guidance throughout the process of transfer to RPI and for undergraduate studies.This project is jointly supported by the Biomolecular Dynamics, Structure and Function Cluster in the Division of Molecular and Cellular Biosciences, the Physics of Living Systems in the Physics Division and the Chemistry of Life Processes program in the Chemistry Division.