SPR Imaging Studies of DNA and Protein Microarrays

点击翻译按钮获取中文摘要

DESCRIPTION (provided by applicant): The overall aim of this research is the development of the label free detection method of surface plasmon resonance (SPR) imaging for the high-throughput study of bioaffinity interactions in an array format. The specific binding interactions of proteins with biomolecular arrays of DNA, peptides, or proteins will be characterized with SPR imaging measurements. A significant portion of this research will focus on the fabrication of well-characterized, highly reproducible, robust, and bioactive arrays of biomolecules on chemically modified gold surfaces. The specific biological systems that will be studied in this project are: (i) the sequence specific binding of bacterial response regulator proteins to DNA arrays in order to help determine how bacteria respond to external stimuli via two component signal transduction systems and gene expression, and to screen libraries of compounds that inhibit the binding of these proteins, (ii) the sequence specific enzymatic activity of kinases, phosphatases and proteases with robust and oriented peptide arrays to help determine how this peptide specificity is used to control cellular processes, and (iii) the design of oriented fusion protein arrays that can be implemented in proteomics studies. The fabrication of oriented fusion protein arrays will require the attachment of capture agents such as glutathione or maltose onto chemically modified gold surfaces in an array format, followed by the binding of fusion proteins that include either glutathione-s-transferase (GST) or maltose binding protein (MBP) onto the array elements. The specific interaction of the GST or MBP with the surface capture agent helps insure that the fusion proteins are oriented in such a manner that they will be accessible to other proteins in solution. In general, the development of SPR imaging techniques for the study of bioaffinity interactions in an array format will help accelerate the search and discovery approaches that are becoming increasingly essential tools of modern biological research.