Biosynthetic studies of proteoglycans in human melanoma cells with a monoclonal antibody to a core glycoprotein of chondroitin sulfate proteoglycans

J Biol Chem. 1984 Oct 25;259(20):12733-41.

Abstract

Monoclonal antibody (Mab) 9.2.27 was utilized in a combination of biosynthetic and biochemical investigations as an immunological probe for the study of chondroitin sulfate proteoglycans (CSP) in human melanoma cells. Pulse-chase and long-term intrinsic labeling immunoprecipitation experiments combined with the biosynthetic inhibitors monensin, cycloheximide, and paranitrophenol-beta-D-xyloside all suggest that Mab 9.2.27 recognizes a set of glycoprotein molecules ranging to a 250-kDa glycoprotein which serves as the core glycoprotein for CSP in human melanoma cells. Peptide maps comparing the 250-kDa and CSP molecule verify that the 250-kDa glycoprotein is the CSP core protein in human melanoma cells. Further studies document that the CSP released by melanoma cells and recognized by Mab 9.2.27 contains (2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-beta-4-O-sulfo-D-galactose and 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-beta-6-O-sulfo-D-galactose saccharides and this CSP can interact with hyaluronic acid-Sepharose. Topographical studies indicate that this CSP has pericellular punctuated distribution on the melanoma cell surface and may play a role in cell-substrate interactions in the biology of metastatic human melanoma.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aggrecans
  • Antibodies, Monoclonal
  • Cell Line
  • Cycloheximide / pharmacology
  • Extracellular Matrix Proteins*
  • Fluorescent Antibody Technique
  • Glycoproteins / biosynthesis*
  • Humans
  • Kinetics
  • Lectins, C-Type
  • Melanoma / metabolism*
  • Neoplasm Metastasis
  • Peptide Fragments / analysis
  • Proteoglycans / biosynthesis*
  • Trypsin

Substances

  • Aggrecans
  • Antibodies, Monoclonal
  • Extracellular Matrix Proteins
  • Glycoproteins
  • Lectins, C-Type
  • Peptide Fragments
  • Proteoglycans
  • Cycloheximide
  • Trypsin