A truncated RHAMM protein for discovering novel therapeutic peptides.
The receptor for hyaluronan mediated motility (RHAMM, gene name HMMR) belongs to a group of proteins that bind to hyaluronan (HA), a high-molecular weight anionic polysaccharide that has pro-angiogenic and inflammatory properties when fragmented. We propose to use a chemically synthesized, truncated version of the protein (706-767), 7 kDa RHAMM, as a target receptor in the screening of novel peptide-based therapeutic agents. Chemical synthesis by Fmoc-based solid-phase peptide synthesis, and optimization using pseudoprolines, results in RHAMM protein of higher purity and yield than synthesis by recombinant protein production. 7 kDa RHAMM was evaluated for its secondary structure, ability to bind the native ligand, HA, and its bioactivity. This 62-amino acid polypeptide replicates the HA binding properties of both native and recombinant RHAMM protein. Furthermore, tubulin-derived HA peptide analogues that bind to recombinant RHAMM and were previously reported to compete with HA for interactions with RHAMM, bind with a similar affinity and specificity to the 7 kDa RHAMM. Therefore, in terms of its key binding properties, the 7 kDa RHAMM mini-protein is a suitable replacement for the full-length recombinant protein.
['Hauser-Kawaguchi|Alexandra|A|', 'Tolg|Cornelia|C|', 'Peart|Teresa|T|', 'Milne|Mark|M|', 'Turley|Eva A|EA|', 'Luyt|Leonard G|LG|']
D002460:Cell Line D004305:Dose-Response Relationship, Drug D016326:Extracellular Matrix Proteins D006801:Humans D018960:Hyaluronan Receptors D015394:Molecular Structure D010455:Peptides D013329:Structure-Activity Relationship
RHAMM Hyaluronan Peptide-based therapeutic agent Pseudoproline Cell adhesion Kinetic evaluation Protein-polysaccharide interaction SPR