Cloning and sequence analysis of cDNA for a neuronal cell membrane antigen, HPC-1.
A monoclonal antibody (mAb), HPC-1, labels the plasma membrane of the amacrine cell soma and inner plexiform layer in rat retina and other central neurons. HPC-1 antigen recognizes several proteins of about 35 kDa. In this study, an HPC-1 positive cDNA, HPC-113, was isolated from a lambda gt11 cDNA library of the rat hippocampus. HPC-113 had the 894-base pair nucleotide sequence in an open reading frame and the calculated molecular mass of the deduced amino acid sequence (298 residues) was 33,989 Da, implying that HPC-113 contains almost the full-length coding region of HPC-1 antigen is an integrated membrane protein revealing the characteristic alpha-helical structure with periodical heptad repeats usually seen in proteins with coiled-coil structures. Although the entire amino acid sequence did not show significant homology to any proteins so far known, a few local sequences in the possible extracellular domain of the HPC-1 antigen molecule had notable homology to some partial sequences in the laminin B1 chain. These sequences of laminin are included in the portion which has neurite outgrowth and/or survival promoting activity. The HPC-1 gene was transcribed in nerve tissues much more predominantly than in non-neuronal tissues. Thus, HPC-1 antigen(s) was confined to be a newly identified neuronal cell membrane protein(s) localized in a subpopulation of neurons.
Inoue
A
A
Department of Physiological Sciences, School of Life Sciences, Graduate University for Advanced Studies, Aichi, Japan.
Obata
K
K
Akagawa
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009474:Neurons / Q000276:immunology*; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D012091:Repetitive Sequences, Nucleic Acid; D012689:Sequence Homology, Nucleic Acid; D050827:Syntaxin 1; D014158:Transcription, Genetic
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C494213:Stx1a protein, rat; D050827:Syntaxin 1; D004247:DNA; D012321:DNA-Directed RNA Polymerases