Chicken CD14, unlike mammalian CD14, is trans-membrane rather than GPI-anchored

Z. Wu, L. Rothwell, T. Hu, P. Kaiser

Research output: Contribution to journalArticlepeer-review

Abstract

A cDNA encoding the chicken homologue of the human myelomonocytic differentiation antigen, CD14, was cloned by RT-PCR from chicken bone marrow cell RNA, using oligonucleotide primers based on the predicted cDNA sequence. The cloned chicken CD14 (chCD14) cDNA encodes an open reading frame of 465 amino acids (aa), with 31-34% aa identity to mouse, bovine and human (hu) CD14. As in mouse and man, chCD14 is a leucine-rich protein. In mammals, CD14 is a GPI-anchored protein. Protein structure analysis suggested that chCD14, by contrast, was potentially a trans-membrane protein. The predicted aa sequence comprises an extracellular domain of 417 aa, followed by a 23-aa trans-membrane segment, and a 25-aa intracytoplasmic region, the latter containing no obvious signalling motifs. COS-7 cells were transfected with p3XFLAG-CMV-8::chCD14 or pCDM8::huCD14, incubated with or without PI-PLC and stained with anti-FLAG or anti-huCD14 antibody respectively. PI-PLC cleaved huCD14 but not chCD14, suggesting that chCD14 is not GPI-anchored. Real-time quantitative RT-PCR analysis revealed that chCD14 mRNA was expressed in most lymphoid and non-lymphoid tissues, except muscle. ChCD14 mRNA was also expressed in most cells examined but strongly expressed in chicken peripheral blood monocyte/macrophages and KUL01+ splenocytes.
Original languageUndefined/Unknown
Pages (from-to)97-104
Number of pages8
JournalDevelopmental and Comparative Immunology
Volume33
Issue number1
DOIs
Publication statusPublished - 2009

Keywords

  • Amino Acid Sequence
  • Animals
  • Antigens, CD14
  • Bone Marrow Cells/metabolism
  • COS Cells
  • Cell Membrane/metabolism
  • Cercopithecus aethiops
  • Chickens
  • DNA, Complementary
  • Humans
  • Molecular Sequence Data
  • Organ Specificity
  • Protein Structure, Tertiary
  • Spleen/cytology
  • Spleen/metabolism

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