Human pancreatic ribonuclease (RNase 1) is a secretory enzyme that can block the flow of biochemical information by catalyzing the cleavage of cellular RNA. Our intent is to use ideas and methods from biological chemistry, molecular biology, and cell biology to reveal key mechanistic aspects of the cytotoxicity of human RNase 1 in physiological and (potentially) clinical settings. During the next grant period, this intent will be achieved in four Specific Aims.
Specific Aims. In Aim 1, we will determine whether the complex of RNase 1 with the cytosolic ribonuclease inhibitor protein (which has femtomolar affinity for RNase 1) acts as a ?sensor? for oxidative damage within a human cell.
In Aim 2, we will determine the structural and physiological roles of the N-linked glycans that are installed on RNase 1.
In Aim 3, we will develop an RNase 1 zymogen as a ?prodrug? that is activated by matrix metalloproteases.
In Aim 4, we will evaluate RNase 1 as an endogenous antimicrobial agent. Significance. The results of the research proposed herein will provide a detailed biochemical understanding of the cytotoxic activity of RNase 1, and could ultimately lead to new chemotherapeutic agents based on an endogenous human enzyme.
This research project is focused on enabling the development of a new class of chemotherapeutic agents based on a protein that is found naturally in the human body. Specifically, the agents are based on ribonuclease, which is an enzyme that can enter human cells and catalyze the cleavage of RNA, leading to cell death. The goal of the project is to obtain fundamental insights into the relationship between the amino-acid sequence of ribonuclease, its three-dimensional structure, and its biological function, and to use those insights to create novel ribonucleases of potential therapeutic utility.
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