Molecular cloning of a glycosylphosphatidylinositol-anchored molecule CDw108.
CDw108, also known as the John-Milton-Hagen human blood group Ag, is an 80-kDa glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that is preferentially expressed on activated lymphocytes and E. The molecular characteristics and biological function of the CDw108 were not clarified previously. In this manuscript, we identify the cDNA clone containing the entire coding sequence of the CDw108 gene and report its molecular characteristics. The 1998-base pairs of the open reading frame of the cloned cDNA encoded a protein of 666 amino acids (aa), including the 46 aa of the signal peptide and the 19 aa of the GPI-anchor motif. Thus, the membrane-anchoring form of CDw108 was the 602 aa, and the estimated molecular mass of the unglycosylated form was 68 kDa. The RGD (Arg-Gly-Asp) cell attachment sequence and the five potential N-linked glycosylation sites were located on the membrane-anchoring form. Flow cytometric and immunoprecipitation analyses of the CDw108 cDNA transfectants confirmed that the cloned cDNA encoded the native form of CDw108. The CDw108 mRNA was expressed in activated PBMCs as well as in the spleen, thymus, testis, placenta, and brain, but was not expressed in any other tissues tested. Radiation hybrid mapping indicated that the CDw108 gene was located in the middle of the long arm of chromosome 15 (15q23-24). This molecular information will be critical for understanding the biological function of the CDw108 Ag.
Yamada
A
A
Cancer Vaccine Development Division, Kurume University Research Center for Innovative Cancer Therapy, Department of Immunology, Japan. akiymd@med.kurume-u.ac.jp
Kubo
K
K
Takeshita
T
T
Harashima
N
N
Kawano
K
K
Mine
T
T
Sagawa
K
K
Sugamura
K
K
Itoh
K
K
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001789:Blood Group Antigens / Q000235:genetics* / Q000276:immunology; D002478:Cells, Cultured; D002874:Chromosome Mapping; D002884:Chromosomes, Human, Pair 15 / Q000276:immunology; D003001:Cloning, Molecular; D058851:GPI-Linked Proteins; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics / Q000276:immunology; D039961:Semaphorins; D014162:Transfection; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D001789:Blood Group Antigens; D058851:GPI-Linked Proteins; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; C118753:SEMA7A protein, human; D039961:Semaphorins