title: Identification, design and synthesis of tubulin-derived peptides as novel hyaluronan mimetic ligands for the receptor for hyaluronan-mediated motility (RHAMM/HMMR).
abstract: Fragments of the extracellular matrix component hyaluronan (HA) promote tissue inflammation, fibrosis and tumor progression. HA fragments act through HA receptors including CD44, LYVE1, TLR2, 4 and the receptor for hyaluronan mediated motility (RHAMM/HMMR). RHAMM is a multifunctional protein with both intracellular and extracellular roles in cell motility and proliferation. Extracellular RHAMM binds directly to HA fragments while intracellular RHAMM binds directly to ERK1 and tubulin. Both HA and regions of tubulin (s-tubulin) are anionic and bind to basic amino acid-rich regions in partner proteins, such as in HA and tubulin binding regions of RHAMM. We used this as a rationale for developing bioinformatics and SPR (surface plasmon resonance) based screening to identify high affinity anionic RHAMM peptide ligands. A library of 12-mer peptides was prepared based on the carboxyl terminal tail sequence of s-tubulin isoforms and assayed for their ability to bind to the HA/tubulin binding region of recombinant RHAMM using SPR. This approach resulted in the isolation of three 12-mer peptides with nanomolar affinity for RHAMM. These peptides bound selectively to RHAMM but not to CD44 or TLR2,4 and blocked RHAMM:HA interactions. Furthermore, fluorescein-peptide uptake by PC3MLN4 prostate cancer cells was blocked by RHAMM mAb but not by CD44 mAb. These peptides also reduced the ability of prostate cancer cells to degrade collagen type I. The selectivity of these novel HA peptide mimics for RHAMM suggest their potential for development as HA mimetic imaging and therapeutic agents for HA-promoted disease.
authors.affiliation: London Regional Cancer Program, London Health Sciences Center, Canada.
authors.forename: Kenneth Virgel N
authors.initials: KV
authors.lastname: Esguerra
authors.forename: Cornelia
authors.initials: C
authors.lastname: Tolg
authors.forename: Natalia
authors.initials: N
authors.lastname: Akentieva
authors.forename: Matthew
authors.initials: M
authors.lastname: Price
authors.forename: Choi-Fong
authors.initials: CF
authors.lastname: Cho
authors.forename: John D
authors.initials: JD
authors.lastname: Lewis
authors.forename: James B
authors.initials: JB
authors.lastname: McCarthy
authors.forename: Eva A
authors.initials: EA
authors.lastname: Turley
authors.forename: Leonard G
authors.initials: LG
authors.lastname: Luyt
mesh_terms: D000595:Amino Acid Sequence; D040761:Biomimetic Materials / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D001943:Breast Neoplasms / Q000378:metabolism; D002232:Carbocyanines; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D004353:Drug Evaluation, Preclinical; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D018960:Hyaluronan Receptors / Q000187:drug effects / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000378:metabolism*; D008024:Ligands; D008297:Male; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness / Q000517:prevention & control; D019151:Peptide Library; D011471:Prostatic Neoplasms / Q000188:drug therapy / Q000378:metabolism; D020349:Surface Plasmon Resonance; D014404:Tubulin / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology
chemical_list: C569686:Alexa Fluor 647; C497483:CD44 protein, human; D002232:Carbocyanines; D005456:Fluorescent Dyes; D018960:Hyaluronan Receptors; D008024:Ligands; D019151:Peptide Library; D014404:Tubulin; D006820:Hyaluronic Acid