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PMID: 1527169 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Homophilic and heterophilic binding activities of Nr-CAM, a nervous system cell adhesion molecule.

The Journal of cell biology ·Vol. 119 ·No. 1 ·1992-10-00 ·Pages 191-202

Mauro VP, Krushel LA, Cunningham BA, Edelman GM

Abstract

Nr-CAM is a membrane glycoprotein that is expressed on neurons. It is structurally related to members of the N-CAM superfamily of neural cell adhesion molecules having six immunoglobulin-like domains and five fibronectin type III repeats in the extracellular region. We have found that the aggregation of chick brain cells was inhibited by anti-Nr-CAM Fab' fragments, indicating that Nr-CAM can act as a cell adhesion molecule. To clarify the mode of action of Nr-CAM, a mouse fibroblast cell line L-M(TK-) (or L cells) was transfected with a DNA expression construct encoding an entire chicken Nr-CAM cDNA sequence. After transfection, L cells expressed Nr-CAM on their surface and aggregated. Aggregation was specifically inhibited by anti-Nr-CAM Fab' fragments. To check the specificity of this aggregation, a fusion protein (FGTNr) consisting of glutathione S-transferase linked to the six immunoglobulin domains and the first fibronectin type III repeat of Nr-CAM was expressed in Escherichia coli. Addition of FGTNr to the transfected cells blocked their aggregation. Further analysis using a combination of cell aggregation assays, binding of cells to FGTNr-coated substrates, aggregation of FGTNr-coated Covaspheres and binding of FGTNr-coated Covaspheres to FGTNr-coated substrates revealed that Nr-CAM mediates two types of cell interactions: a homophilic, divalent cation-independent binding, and a heterophilic, divalent cation-dependent binding. Homophilic binding was demonstrated between transfected L cells, between chick embryo brain cells and FGTNr, and between Covaspheres to which FGTNr was covalently attached. Heterophilic binding was shown to occur between transfected and untransfected L cells, and between FGTNr and primary chick embryo fibroblasts; in all cases, it was dependent on the presence of either calcium or magnesium. Primary chick embryo glia or a human glial cell line did not bind to FGTNr-coated substrates. The results indicate that Nr-CAM is a cell adhesion molecule of the nervous system that can bind by two distinct mechanisms, a homophilic mechanism that can mediate interactions between neurons and a heterophilic mechanism that can mediate binding between neurons and other cells such as fibroblasts.

MeSH Terms
Animals Avian Proteins Cations, Divalent/metabolism Cell Adhesion/physiology Cell Adhesion Molecules Cell Adhesion Molecules, Neuronal/metabolism Chick Embryo Cloning, Molecular Fluorescent Antibody Technique Fluorescent Dyes L Cells Mice Protein Conformation Recombinant Fusion Proteins/metabolism Transfection
Chemicals
Avian Proteins Cations, Divalent Cell Adhesion Molecules Cell Adhesion Molecules, Neuronal Fluorescent Dyes NRCAM protein, Gallus gallus NRCAM protein, human Nrcam protein, mouse Recombinant Fusion Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Mauro V P
Rockefeller University, New York 10021.
Krushel L A
Cunningham B A
Edelman G M
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.119.1.191
Published
1992-10-00
Pages
191-202
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289630
Subset
IM
Grants
NICHD NIH HHS · HD-09635 · United States
NICHD NIH HHS · HD16550 · United States
NINDS NIH HHS · NS-28932 · United States
External Links
PubMed source DOI: 10.1083/jcb.119.1.191 PMC Full Text View journal details
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