Abstract
The human homologue of 4-1BB (H4-1BB) cDNA was isolated from PMA plus ionomycin-treated human peripheral T-cell cDNA libraries. The amino acid sequence deduced from the nucleotide sequence showed that the protein is composed of 255 amino acids with 2 potential N-linked glycosylation sites. The molecular weight of its protein backbone is calculated to be 27 kDa. The H4-1BB contains features such as signal sequence and transmembrane domain, indicating that it is a receptor protein. This protein showed 60% identity of amino acid sequence to mouse 4-1BB. In the cytoplasmic domain there are 5 regions of amino acid sequences conserved from mouse to human, indicating that these residues might be important in the 4-1BB function. H4-1BB mRNA was detected in unstimulated peripheral blood T cells and was inducible in T-cell lines such as Jurkat and CEM. H4-1BB-AP, a fusion protein between the H4-1BB extracellular domain and alkaline phosphatase, was used to identify the ligand for the H4-1BB. Although the H4-1BB ligand was detected in both T and B cells of human peripheral blood, the ligand was preferentially expressed in primary B cells and B-cell lines. Daudi, a B-cell lymphoma, was one of the B-cell lines that carried a higher number of ligands. Scatchard analysis showed that the Kd = 1.4 x 10(9) M and the number of ligands in Daudi cell was 4.2 x 10(3).
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3T3 Cells
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4-1BB Ligand
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Amino Acid Sequence
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Animals
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Antigens, CD
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B-Lymphocytes / metabolism
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Base Sequence
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DNA, Complementary / genetics
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Humans
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Leukemia-Lymphoma, Adult T-Cell / pathology
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Lymphocyte Activation
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Membrane Glycoproteins / chemistry*
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / isolation & purification
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Mice
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Molecular Sequence Data
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Molecular Weight
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Polymerase Chain Reaction
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Precursor Cell Lymphoblastic Leukemia-Lymphoma / pathology
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Protein Structure, Tertiary
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RNA, Messenger / analysis
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Receptors, Nerve Growth Factor / chemistry*
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Receptors, Nerve Growth Factor / genetics
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Receptors, Nerve Growth Factor / isolation & purification
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Receptors, Tumor Necrosis Factor / chemistry*
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Receptors, Tumor Necrosis Factor / genetics
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Receptors, Tumor Necrosis Factor / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Species Specificity
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T-Lymphocytes / drug effects
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T-Lymphocytes / metabolism
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Tumor Cells, Cultured
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Tumor Necrosis Factor Receptor Superfamily, Member 9
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Tumor Necrosis Factor-alpha / chemistry*
Substances
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4-1BB Ligand
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Antigens, CD
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DNA, Complementary
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Membrane Glycoproteins
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Peptide Fragments
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RNA, Messenger
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Receptors, Nerve Growth Factor
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Receptors, Tumor Necrosis Factor
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Recombinant Fusion Proteins
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TNFRSF9 protein, human
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TNFSF9 protein, human
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Tnfrsf9 protein, mouse
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Tnfsf9 protein, mouse
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Tumor Necrosis Factor Receptor Superfamily, Member 9
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Tumor Necrosis Factor-alpha