A truncated RHAMM protein for discovering novel therapeutic peptides

Alexandra Hauser-Kawaguchi; Cornelia Tolg; Teresa Peart; Mark Milne; Eva A Turley; Leonard G Luyt

doi:10.1016/j.bmc.2018.09.018

A truncated RHAMM protein for discovering novel therapeutic peptides

Bioorg Med Chem. 2018 Oct 1;26(18):5194-5203. doi: 10.1016/j.bmc.2018.09.018. Epub 2018 Sep 18.

Authors

Alexandra Hauser-Kawaguchi¹, Cornelia Tolg², Teresa Peart², Mark Milne², Eva A Turley³, Leonard G Luyt⁴

Affiliations

¹ Department of Chemistry, Western University, London, Ontario, Canada.
² Cancer Research Laboratory Program, Lawson Health Research Institute and London Regional Cancer Program, London Health Sciences Center, London, Ontario, Canada.
³ Cancer Research Laboratory Program, Lawson Health Research Institute and London Regional Cancer Program, London Health Sciences Center, London, Ontario, Canada; Department of Biochemistry, Western University, London, Ontario, Canada; Department of Oncology, Western University, London, Ontario, Canada.
⁴ Department of Chemistry, Western University, London, Ontario, Canada; Cancer Research Laboratory Program, Lawson Health Research Institute and London Regional Cancer Program, London Health Sciences Center, London, Ontario, Canada; Department of Oncology, Western University, London, Ontario, Canada; Department of Medical Imaging, Western University, London, Ontario, Canada. Electronic address: lluyt@uwo.ca.

PMID: 30249497
DOI: 10.1016/j.bmc.2018.09.018

Abstract

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.

Keywords: Cell adhesion; Hyaluronan; Kinetic evaluation; Peptide-based therapeutic agent; Protein-polysaccharide interaction; Pseudoproline; RHAMM; SPR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Dose-Response Relationship, Drug
Extracellular Matrix Proteins / antagonists & inhibitors*
Extracellular Matrix Proteins / metabolism
Humans
Hyaluronan Receptors / antagonists & inhibitors*
Hyaluronan Receptors / metabolism
Molecular Structure
Peptides / chemical synthesis
Peptides / chemistry
Peptides / pharmacology*
Structure-Activity Relationship

Substances

Extracellular Matrix Proteins
Hyaluronan Receptors
Peptides
hyaluronan-mediated motility receptor