A role of chondroitin sulfate glycosaminoglycan binding site in alpha4beta1 integrin-mediated melanoma cell adhesion.
We have previously reported that alpha4beta1 (but not alpha5beta1) integrin-mediated melanoma cell adhesion is inhibited by removal of cell surface chondroitin sulfate glycosaminoglycan (CSGAG), suggesting that melanoma chondroitin sulfate proteoglycan plays a role in modulating the adhesive function of alpha4beta1 integrin. In the current study, we demonstrated that alpha4beta1 integrin binds to CSGAG. We have identified a peptide from within alpha4 integrin termed SG1 (KKEKDIMKKTI) that binds to cell surface melanoma chondroitin sulfate proteoglycan, indicating that SG1 represents a CSGAG binding site within the alpha4 integrin subunit. Soluble SG1 inhibits alpha4beta1 integrin-mediated human melanoma cell adhesion to CS1. Polyclonal antibody generated against the peptide inhibits melanoma cell adhesion to CS1, and the inhibition is reversed by Mn2+ and an activating monoclonal antibody anti-beta1 (8A2). Additionally, pretreatment of cells with anti-SG1 IgG inhibits the expression of the monoclonal antibody 15/7 epitope in the presence of soluble CS1 peptide, suggesting that anti-SG1 IgG prevents ligand binding by alpha4beta1 integrin. These results demonstrate that alpha4beta1 integrin interacts directly with CSGAG through SG1 site, and that this site can affect the ligand binding properties of the integrin.
Iida
J
J
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Meijne
A M
AM
Oegema
T R
TR
Yednock
T A
TA
Kovach
N L
NL
Furcht
L T
LT
McCarthy
J B
JB
D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D001665:Binding Sites / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D002809:Chondroitin Sulfates / Q000502:physiology*; D006801:Humans; D039041:Integrin alpha4beta1; D016023:Integrins / Q000737:chemistry* / Q000502:physiology; D008345:Manganese / Q000494:pharmacology; D008545:Melanoma / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry; D011485:Protein Binding / Q000502:physiology; D011509:Proteoglycans / Q000502:physiology*; D016201:Receptors, Lymphocyte Homing / Q000737:chemistry* / Q000502:physiology; D014407:Tumor Cells, Cultured
D000906:Antibodies; D039041:Integrin alpha4beta1; D016023:Integrins; D010446:Peptide Fragments; D011509:Proteoglycans; D016201:Receptors, Lymphocyte Homing; D008345:Manganese; D002809:Chondroitin Sulfates