Molecular cloning of cDNA encoding MS2 antigen, a novel cell surface antigen strongly expressed in murine monocytic lineage.
cDNA clones which strongly hybridized with a 3.1 kb mRNA from mouse macrophages and macrophage cell lines and weakly with mRNA from P815 but not from a variety of other cell lines and tissues were isolated from cDNA libraries constructed using mRNA from murine macrophage cell lines and peritoneal macrophages. Treatment of a macrophage cell line with macrophage stimulators significantly enhanced transcription of the mRNA. Sequencing analysis of these clones demonstrated that the cDNA consisted of 3036 bp insert containing a 2478 bp open reading frame followed by a 538 bp 3' untranslated region. The amino acid sequence, deduced from the nucleotide sequence of the cDNA, predicted a protein containing a signal peptide, an extracellular region, a transmembrane domain, and a cytoplasmic tail. The extracellular region had five putative N-glycosylation sites and a cysteine-rich domain, whereas the cytoplasmic region consisted of a proline-rich amino acid sequence significantly similar to CD2. SDS-PAGE and NEPHGE SDS-PAGE analysis of the immunoprecipitated membrane of the macrophage cell lines prepared by using rabbit anti-MS2 peptide antibody raised against a synthetic peptide preparation relative to a hydrophilic region of the MS2 amino acid sequence confirmed that MS2 protein is a cross-linked protein having approximate molecular sizes of 89 kd and pl 6.5-7.0. These results show that MS2 protein is a novel cell surface antigen expressed mainly in monocytic lineages.
Yoshida
S
S
Department of Pathology, Medical College of Oita, Japan.
Setoguchi
M
M
Higuchi
Y
Y
Akizuki
S
S
Yamamoto
S
S
D051722:ADAM Proteins; D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics; D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D018801:CD2 Antigens; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D007526:Isoelectric Point; D008565:Membrane Proteins; D008666:Metalloendopeptidases; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009000:Monocytes / Q000276:immunology*; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics; D011971:Receptors, Immunologic / Q000235:genetics; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D000954:Antigens, Surface; D018801:CD2 Antigens; D008565:Membrane Proteins; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D004247:DNA; D051722:ADAM Proteins; C489402:Adam8 protein, mouse; D008666:Metalloendopeptidases