Elastin peptides signaling relies on neuraminidase-1-dependent lactosylceramide generation

PLoS One. 2010 Nov 16;5(11):e14010. doi: 10.1371/journal.pone.0014010.

Abstract

The sialidase activity of neuraminidase-1 (Neu-1) is responsible for ERK 1/2 pathway activation following binding of elastin peptide on the elastin receptor complex. In this work, we demonstrate that the receptor and lipid rafts colocalize at the plasma membrane. We also show that the disruption of these microdomains as well as their depletion in glycolipids blocks the receptor signaling. Following elastin peptide treatment, the cellular GM(3) level decreases while lactosylceramide (LacCer) content increases consistently with a GM(3)/LacCer conversion. The use of lactose or Neu-1 siRNA blocks this process suggesting that the elastin receptor complex is responsible for this lipid conversion. Flow cytometry analysis confirms this elastin peptide-driven LacCer generation. Further, the use of a monoclonal anti-GM(3) blocking antibody shows that GM(3) is required for signaling. In conclusion, our data strongly suggest that Neu-1-dependent GM(3)/LacCer conversion is the key event leading to signaling by the elastin receptor complex. As a consequence, we propose that LacCer is an early messenger for this receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antigens, CD / biosynthesis*
  • Antigens, CD / metabolism
  • Blotting, Western
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Elastin / chemistry
  • Elastin / pharmacology*
  • Enzyme Activation / drug effects
  • Fibroblasts / cytology
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Flow Cytometry
  • G(M3) Ganglioside / metabolism
  • Humans
  • Lactose / pharmacology
  • Lactosylceramides / biosynthesis*
  • Lactosylceramides / metabolism
  • Membrane Microdomains / metabolism
  • Middle Aged
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Neuraminidase / genetics
  • Neuraminidase / metabolism*
  • Peptides / pharmacology*
  • RNA Interference
  • Receptors, Cell Surface / antagonists & inhibitors
  • Receptors, Cell Surface / metabolism
  • Signal Transduction
  • Young Adult

Substances

  • Antigens, CD
  • G(M3) Ganglioside
  • Lactosylceramides
  • Peptides
  • Receptors, Cell Surface
  • elastin-binding proteins
  • CDw17 antigen
  • Elastin
  • MAPK1 protein, human
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Neuraminidase
  • Lactose