University of Colorado Department of Biochemistry and Molecular Genetics

​Synthetic peptide and antipeptide approaches play major roles in understanding protein structure and function. Our research interests range from:

1. The de novo design of model proteins, an important endeavor that not only tests our understanding of protein folding and structure but also lays the groundwork for the design of novel proteins with the desired biological/immunological activities. We have selected the two-stranded a-helical coiled-coil as an ideal model system for studying protein stability, subunit interactions and protein folding. In addition, we are studying native coiled-coils in tropomyosin, the transporter protein ProP and in the motor proteins, kinesin and kinesin-like proteins, dynein and dynamin;
   

2. Our expertise in understanding the formation and stability of coiled-coils has resulted in a new project to investigate the coiled-coil domains in the SARS (Severe Acute Respiratory Syndrome) coronavirus spike glycoprotein. The aim is to design and synthesize stable peptides and peptidomimetics that will inhibit SARS virus infection of human cells;

3. The design and engineering of peptides for biotechnology applications. This area includes the development of novel antimicrobial cyclic peptides with enhanced antimicrobial activity, low toxicity with broad spectrum activity or activity profiles for selected clinical indications. The development of a novel peptidylmimetic as a therapeutic to treat Pseudomonas aeruginosa. The design of comformationally defined and constrained combinatorial peptide libraries for drug discovery. The de novo design of templates to prepare antibodies that recognize a-helical protein sequences in native proteins;
   

4. The development of synthetic peptide anti-adhesin bacterial vaccines and antibody therapeutics. These antigen-antibody studies will also investigate such issues as strain-specificity, cross-reactivity, and immunogen presentation as a peptide versus the same region presented in the native protein, which will lead to a better understanding of the immune response at the molecular level;
   

5. The development of novel HPLC and capillary electrophoretic methods for the separation of peptides and proteins for proteomic applications. Our goal is to replace two-dimensional gel electrophoresis with two-dimensional chromatography or a combined chromatography/capillary electrophoresis method prior to MS/MS. In addition, we will continue our studies to demonstrate the general feasibility of equating polypeptide reversed-phase chromatography (RPC) elution behavior with peptide and protein stability, conformation and function. Of particular significance is our recent introduction of "temperature profiling" in RPC which enables a highly sensitive monitoring of the association of peptide molecules either through oligomerization or aggregation;
   

6. The above projects involve a wide range of technologies in protein chemistry: peptide synthesis, HPLC, CE, amino acid analysis, mass spectrometry and characterization of peptides and proteins by CD and NMR spectroscopy, analytical ultracentrifugation, Biacore analysis and a range of bioassays.