The NMR facility of the LMC has two main functions: The first is as a resource for the entire chemist community of the LMC and NCI Frederick to use as a routine use facility to walk up and collect NMR data on synthetic compounds prepared for projects under the auspices of other PI's and 2) It is used to tackle more challenging problems related to the structure and 2-dimensional conformations of drug molecules or other biopolymers (peptide, glycopeptides, oligonucleotides and oligosaccharides) studied in the various sections. This project relates to number (2) in that I use part of my time to do NMR-based conformational analysis that is a critical part of my work and of the entire LMC. Thus the creation of this new project is to define the effort dedicated to original research in the area of conformational analysis of molecules of biological significance to the LMC and NCI as a whole. In the past fiscal year, we have de-emphasized the study of DNA containing """"""""locked"""""""" nucleosides which we have already spent considerable effort and have concentrated on the study of interesting peptides and glycopeptides in various solutions. We have defined the solution conformation of an integrin binding peptide of thrombospondin-1, an important extracellular matrix protein with a wealth of biological functions. This peptide can promote or inhibit angiogenesis and it contains a 3-amino acid motif that is critical for its activity. We have defined the conformation of this motif in water and in dodecylphosphocholine micelles. The conformations were compared to a newly derived crystal structure and we now are in a position to design non-peptide molecules to mimic this peptide and possibly as drug candidates.In a separte project, we have defined the solution conformation of a very important small glycopeptide called antiproliferative factor (APF, from our PNAS paper of 2004) which is the causative agent of interstitial cystitis, a painful inflammatory disease of the bladder. APF also inhibits bladder tumor cell rowth at sub-nanomolar concentrations. The peptide is virtually unstructured in water solution but assumes some structure in membrane mimicking environments (trifluoroethanol). This work will continue with the study of the structures of other analogues of APF synthesized in the laboratory of Christopher Michejda. Other work involves the construction of a database of NMR data for the nucleoside analogues prepared in the LMC and answering basic science questions relating to the structures of the interesting locked nucleoside analogues prepared in the Section of Victor Marquez. We will continue to study the DNA molecules but on a reduced effort basis. We are actively soliciting the aid of other labs to assist in this project since it is a highly important area of research in both our lab and in many labs around the world.
