Melanoma cell CD44 interaction with the alpha 1(IV)1263-1277 region from basement membrane collagen is modulated by ligand glycosylation.

Abstract

Invasion of the basement membrane is believed to be a critical step in the metastatic process. Melanoma cells have been shown previously to bind distinct triple-helical regions within basement membrane (type IV) collagen. Additionally, tumor cell binding sites within type IV collagen contain glycosylated hydroxylysine residues. In the present study, we have utilized triple-helical models of the type IV collagen alpha1(IV)1263-1277 sequence to (a) determine the melanoma cell receptor for this ligand and (b) analyze the results of single-site glycosylation on melanoma cell recognition. Receptor identification was achieved by a combination of methods, including (a) cell adhesion and spreading assays using triple-helical alpha1(IV)1263-1277 and an Asp(1266)Abu variant, (b) inhibition of cell adhesion and spreading assays, and (c) triple-helical alpha1(IV)1263-1277 affinity chromatography with whole cell lysates and glycosaminoglycans. Triple-helical alpha1(IV)1263-1277 was bound by melanoma cell CD44/chondroitin sulfate proteoglycan receptors and not by the collagen-binding integrins or melanoma-associated proteoglycan. Melanoma cell adhesion to and spreading on the triple-helical alpha1(IV)1263-1277 sequence was then compared for glycosylated (replacement of Lys(1265) with Hyl(O-beta-d-galactopyranosyl)) versus non-glycosylated ligand. Glycosylation was found to strongly modulate both activities, as adhesion and spreading were dramatically decreased due to the presence of galactose. CD44/chondroitin sulfate proteoglycan did not bind to glycosylated alpha1(IV)1263-1277. Overall, this study (a) is the first demonstration of the prophylactic effects of glycosylation on tumor cell interaction with the basement membrane, (b) provides a rare example of an apparent unfavorable interaction between carbohydrates, and (c) suggests that sugars may mask "cryptic sites" accessible to tumor cells with cell surface or secreted glycosidase activities.

• Authors: Janelle L Lauer-Fields, Navdeep B Malkar, Gérard Richet, Karlheinz Drauz, Gregg B Fields
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 278 Issue 16 Pg. 14321-30 (Apr 18 2003) ISSN: 0021-9258 [Print] United States
• PMID: 12574156 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Amino Acids
• Carbazoles
• Chondroitin Sulfate Proteoglycans
• Collagen Type IV
• Glycosaminoglycans
• Hyaluronan Receptors
• Ligands
• Hydroxylysine
• Aspartic Acid
• Sodium Chloride
• Lysine

Topics

• Amino Acids (chemistry)
• Aspartic Acid (chemistry)
• Binding Sites
• Carbazoles (chemistry)
• Cell Adhesion
• Chondroitin Sulfate Proteoglycans (metabolism)
• Chromatography, High Pressure Liquid
• Circular Dichroism
• Collagen Type IV (metabolism)
• Dose-Response Relationship, Drug
• Enzyme-Linked Immunosorbent Assay
• Glycosaminoglycans (metabolism)
• Glycosylation
• Humans
• Hyaluronan Receptors (metabolism)
• Hydroxylysine (metabolism)
• Ligands
• Lysine (chemistry)
• Melanoma (metabolism)
• Peptide Biosynthesis
• Precipitin Tests
• Protein Binding
• Sodium Chloride (pharmacology)
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• Temperature
• Tumor Cells, Cultured