Characterization of densin-180, a new brain-specific synaptic protein of the O-sialoglycoprotein family.
We purified an abundant protein of apparent molecular mass 180 kDa from the postsynaptic density fraction of rat forebrain and obtained amino acid sequences of three tryptic peptides generated from the protein. The sequences were used to design a strategy for cloning the cDNA encoding the protein by polymerase chain reaction. The open reading frame of the cDNA encodes a novel protein of predicted molecular mass 167 kDa. We have named the protein densin-180. Antibodies raised against the predicted amino and carboxyl sequences of densin-180 recognize a 180 kDa band on immunoblots that is enriched in the postsynaptic density fraction. Immunocytochemical localization of densin-180 in dissociated hippocampal neuronal cultures shows that the protein is highly concentrated at synapses along dendrites. The message encoding densin-180 is brain specific and is more abundant in forebrain than in cerebellum. The sequence of densin-180 contains 17 leucine-rich repeats, a sialomucin domain, an apparent transmembrane domain, and a PDZ domain. This arrangement of domains is similar to that of several adhesion molecules, in particular GPIbalpha, which mediates binding of platelets to von Willebrand factor. We propose that densin-180 participates in specific adhesion between presynaptic and postsynaptic membranes at glutamatergic synapses.
Apperson
M L
ML
Division of Biology, California Institute of Technology, Pasadena, California 91125, USA.
Moon
I S
IS
Kennedy
M B
MB
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry / Q000235:genetics*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured / Q000737:chemistry / Q000648:ultrastructure; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000235:genetics; D006624:Hippocampus / Q000737:chemistry / Q000166:cytology; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009474:Neurons / Q000737:chemistry* / Q000648:ultrastructure; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D017729:Presynaptic Terminals / Q000737:chemistry / Q000201:enzymology; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D012795:Sialoglycoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013569:Synapses / Q000737:chemistry / Q000201:enzymology
D015815:Cell Adhesion Molecules; D018076:DNA, Complementary; C504463:Lrrc7 protein, rat; D008565:Membrane Proteins; D012795:Sialoglycoproteins; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases