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PMID: 2045418 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Structure of a new nervous system glycoprotein, Nr-CAM, and its relationship to subgroups of neural cell adhesion molecules.

The Journal of cell biology ·Vol. 113 ·No. 6 ·1991-06-00 ·Pages 1399-412

Grumet M, Mauro V, Burgoon MP, Edelman GM, Cunningham BA

Abstract

We have identified and characterized a new glycoprotein in the chicken nervous system using immunological and molecular biological methods and we have examined its tissue distribution. Analysis revealed that this protein is very similar in structure to the chicken neuron-glia cell adhesion molecule, Ng-CAM, and to mouse L1. cDNA clones encompassing the entire coding sequence of this Ng-CAM related molecule, called Nr-CAM, have been isolated and sequenced. A glycoprotein containing one major component of Mr 145,000 on SDS-PAGE was purified from brain by lentil lectin affinity chromatography and FPLC, and its amino-terminal sequence was identical to that predicted from the Nr-CAM cDNA. The complete cDNA sequence encodes six Ig-like domains, five fibronectin type III repeats, a predicted transmembrane domain, and a short cytoplasmic domain. On Northern blots, nucleic acid probes for Nr-CAM recognized one major RNA species of approximately 7 kb and much lesser amounts of larger RNAs. Most of the same probes hybridized to single bands on genomic Southern blots, suggesting that Nr-CAM is encoded by a single gene that may be alternatively processed to yield several mRNAs. In support of this notion, two Nr-CAM cDNA clones had a 57-bp sequence located between the second and third Ig-like domains that was not found in two other Nr-CAM cDNA clones, and two other clones were isolated that lacked the 279-bp segment encoding the fifth fibronectin-like type III repeat. Antibodies against the purified protein and synthetic peptides in Nr-CAM both recognized a predominant Mr 145,000 species and a much less prevalent species of Mr 170,000 in neural tissues. Levels of Nr-CAM expression increased in the brain until approximately embryonic day (E) 12, followed by slightly lower levels of expression at E18 and after hatching. Immunofluorescent staining with anti-Nr-CAM antibodies showed that most neurons in the retina were positive at E7 and the pattern of expression became restricted to several layers on neuronal cell bodies and fibers during development. Anti-Nr-CAM antibodies labeled specifically cell surfaces on neurons in culture. Although the structure of Nr-CAM resembles that of chicken Ng-CAM and mouse L1, the identity with each of these neural CAMs does not exceed 40%. The differences indicate that Nr-CAM is distinct from Ng-CAM and L1, but there are sufficient similarities to suggest that all of these molecules are members of a subgroup of neural CAMs in the N-CAM superfamily.(ABSTRACT TRUNCATED AT 400 WORDS)

MeSH Terms
Amino Acid Sequence Animals Avian Proteins Base Sequence Blotting, Northern Blotting, Southern Brain Chemistry Cell Adhesion Molecules Cell Adhesion Molecules, Neuronal/analysis,genetics Chick Embryo Cloning, Molecular DNA/isolation & purification Fluorescent Antibody Technique Mice Molecular Sequence Data Retina/chemistry Sequence Alignment
Chemicals
Avian Proteins Cell Adhesion Molecules Cell Adhesion Molecules, Neuronal NRCAM protein, Gallus gallus Nrcam protein, mouse DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Grumet M
Rockefeller University, New York, New York 10021.
Mauro V
Burgoon M P
Edelman G M
Cunningham B A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.113.6.1399
Published
1991-06-00
Pages
1399-412
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289038
Subset
IM
Grants
NICHD NIH HHS · HD-09635 · United States
NINDS NIH HHS · NS-21629 · United States
NINDS NIH HHS · NS-28932 · United States
Databases
GENBANK
X58482
External Links
PubMed source DOI: 10.1083/jcb.113.6.1399 PMC Full Text View journal details
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