Evidence for direct amelogenin-target cell interactions using dynamic force spectroscopy

Research output: Contribution to journalArticlepeer-review

Authors

  • Jennifer Kirkham
  • Colin Robinson
  • Steven J Brookes
  • Roger C Shore
  • D Alastair Smith

Colleges, School and Institutes

Abstract

Increasing evidence suggests that amelogenin, long held to be a structural protein of developing enamel matrix, may also have cell signaling functions. However, a mechanism for amelogenin cell signaling has yet to be described. The aim of the present study was to use dynamic chemical force spectroscopy to measure amelogenin interactions with possible target cells. Full-length amelogenin (rM179) was covalently attached to silicon nitride AFM tips. Synthetic RGD peptides and unmodified AFM tips were used as controls. Amelogenin-RGD cell binding force measurements were carried out using human periodontal ligament fibroblasts (HPDF) from primary explants and a commercially available osteoblast-like human sarcoma cell line as the targets. Results indicated a linear logarithmic dependence between loading rate and unbinding force for amelogenin-RGD target cells across the range of loading rates used. For RGD controls, binding events measured at 5.5 nN s-1 force loading rate resulted in a mean force of 60 pN. Values for amelogenin-fibroblast and amelogenin-osteoblast-like cell unbinding forces, measured at similar loading rates, were 50 and 55 pN, respectively. These data suggest that amelogenin interacts with potential target cells with forces characteristic of specific ligand-receptor binding, suggesting a direct effect for amelogenin at target cell membranes.

Details

Original languageEnglish
Pages (from-to)219-24; discussion 254-6, 381-2
JournalEuropean Journal of Oral Sciences
Volume114 Suppl 1
Publication statusPublished - May 2006

Keywords

  • Amelogenin, Cell Communication, Cell Line, Tumor, Cells, Cultured, Dental Enamel Proteins, Fibroblasts, Humans, Intercellular Signaling Peptides and Proteins, Lipid Bilayers, Microscopy, Atomic Force, Oligopeptides, Osteoblasts, Periodontal Ligament, Receptors, Cell Surface, Signal Transduction, Silicon Compounds, Spectrum Analysis