Cloning, recombinant expression and biochemical characterization of the murine CD83 molecule which is specifically upregulated during dendritic cell maturation

• Published in: FEBS letters 1999-11, Vol.461 (3), p.211-216
• Main Authors: Berchtold, Susanne, Mühl-Zürbes, Petra, Heufler, Christine, Winklehner, Patrizia, Schuler, Gerold, Steinkasserer, Alexander
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antigens, CD; Base Sequence; CD83; CD83 Antigen; cDNA; Cell Differentiation; Cloning, Molecular; COS Cells; DC, dendritic cells; Dendritic cell; Dendritic Cells - cytology; Dendritic Cells - metabolism; DNA, Complementary - genetics; Escherichia coli; Gene Expression Regulation - drug effects; Glycosylation; GM-CSF, granulocyte-macrophage colony-stimulating factor; Humans; Immunoglobulins - biosynthesis; Immunoglobulins - chemistry; Immunoglobulins - genetics; Lipopolysaccharides - pharmacology; LPS, lipopolysaccharide; Magnetic Resonance Spectroscopy; Membrane Glycoproteins - biosynthesis; Membrane Glycoproteins - chemistry; Membrane Glycoproteins - genetics; Mice; Molecular Sequence Data; NMR, nuclear magnetic resonance; PCR, polymerase chain reaction; Protein Conformation; Protein Processing, Post-Translational; Protein Sorting Signals - chemistry; Protein Structure, Tertiary; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant protein; RNA, Messenger - biosynthesis; TNF-α, tumor necrosis factor α; Tumor Necrosis Factor-alpha - pharmacology
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(99)01465-9

Human CD83 (hCD83) is a glycoprotein expressed predominantly on the surface of dendritic cells (DC) and represents the best marker for mature DC. Here, we report the cloning of the cDNA encoding mouse CD83 (mCD83) from a murine bone marrow-derived DC (BM-DC) cDNA library. DNA sequence analysis revealed a 196 amino acid protein including a signal peptide of 21 amino acids which shares 63% amino acid identity with hCD83. Using Northern blot analyses, mCD83 mRNA was found to be strongly expressed in mouse BM-DC and its expression was upregulated following stimulation with LPS or TNF-α. Transfection experiments using COS-7 cells revealed that mCD83 is glycosylated. Furthermore, the extracellular CD83 domain was recombinantly expressed in Escherichia coli and one-dimensional NMR data strongly support that the protein is structurally folded.