Identification of mammalian and invertebrate analogues of the avian calcium-dependent cell adhesion protein N-cadherin with synthetic-peptide directed antibodies against a conserved cytoplasmic domain.
N-cadherin, a 130kD transmembrane adhesive glycoprotein, is a mediator of specific cellular interactions during development. Analysis of N-cadherin at the protein level, to date, has been largely dependent upon monoclonal antibody NCD-2 which recognizes only avian N-cadherin. We produced a monospecific polyclonal antiserum, C-NCAD(838-856), to a synthetic peptide corresponding to a portion of the highly conserved c-terminal cytoplasmic domain of chick N-cadherin. Using polyacrylamide gel electrophoresis and immunoblotting to map tissue distribution we show that the antiserum detects chick N-cadherin with a similar tissue distribution as NCD-2. Unlike NCD-2, however, anti-C-NCAD(838-856) recognizes N-cadherin analogues in a wide variety of species, including mouse, human, fish and drosophila. The results of comparative immunoblot studies demonstrate similar tissue-specific patterns and apparent molecular weight variation in the chick, mouse and human. This indicates that N-cadherin structure and expression, and most likely function as well, have been highly conserved in evolution. The antiserum recognizes an epitope unique to N-cadherin which is conserved among N-cadherins from a variety of species but is absent from other members of the cadherin gene family, as no immunoreactivity was detected with tissues bearing these other cadherins. The antiserum is thus a useful tool for the phylogenetic and biochemical investigation of N-cadherin from a variety of tissue sources.
Lagunowich
L A
LA
Department of Anatomy, Thomas Jefferson University, Philadelphia, PA 19107.
Donoso
L A
LA
Grunwald
G B
GB
D000595:Amino Acid Sequence; D000811:Anguilla; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D015820:Cadherins / Q000032:analysis* / Q000276:immunology; D002642:Chick Embryo; D002645:Chickens; D004331:Drosophila melanogaster / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005123:Eye / Q000737:chemistry; D005260:Female; D005849:Gerbillinae; D006801:Humans; D006829:Hydra; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis; D013045:Species Specificity; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D015820:Cadherins; D010455:Peptides