Structural basis for inflammation-driven shedding of CD163 ectodomain and tumor necrosis factor-α in macrophages

Anders Etzerodt, Mie Rostved Rasmussen, Pia Svendsen, Athena Chalaris, Jeanette Schwarz, Ian Galea, Holger Jon Møller, Søren Kragh Moestrup

Research output: Contribution to journalJournal articleResearchpeer-review


The haptoglobin-hemoglobin receptor CD163 and proTNF-α are transmembrane macrophage proteins subjected to cleavage by the inflammation-responsive protease ADAM17. This leads to release of soluble CD163 (sCD163) and bioactive TNF-α. Sequence comparison of the juxtamembrane region identified similar palindromic sequences in human CD163 ((1044)Arg-Ser-Ser-Arg) and proTNF-α ((78)Arg-Ser-Ser-Ser-Arg). In proTNF-α the Arg-Ser-Ser-Ser-Arg sequence is situated next to the previously established ADAM17 cleavage site. Site-directed mutagenesis revealed that the sequences harbor essential information for efficient cleavage of the two proteins upon ADAM17 stimulation. This was further evidenced by analysis of mouse CD163 that, like CD163 in other non-primates, does not contain the palindromic CD163 sequence in the juxtamembrane region. Mouse CD163 resisted endotoxin- and phorbol ester-induced shedding, and ex vivo analysis of knock-in of the Arg-Ser-Ser-Arg sequence in mouse CD163 revealed a receptor shedding comparable with that of human CD163. In conclusion, we have identified an essential substrate motif for ADAM17-mediated CD163 and proTNF-α cleavage in macrophages. In addition, the present data indicate that CD163, by incorporation of this motif in late evolution, underwent a modification that allows for an instant down-regulation of surface CD163 expression and inhibition of hemoglobin uptake. This regulatory modality seems to have coincided with the evolution of an enhanced hemoglobin-protecting role of the haptoglobin-CD163 system in primates.

Original languageEnglish
JournalJournal of Biological Chemistry
Issue number2
Pages (from-to)778-788
Publication statusPublished - 10. Jan 2014
Externally publishedYes


  • ADAM Proteins
  • Amino Acid Sequence
  • Animals
  • Antigens, CD
  • Antigens, Differentiation, Myelomonocytic
  • Binding Sites
  • Cells, Cultured
  • Endotoxemia
  • Enzyme Precursors
  • HEK293 Cells
  • Humans
  • Immunoblotting
  • Inflammation
  • Lipopolysaccharides
  • Macrophages
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Molecular Sequence Data
  • Mutation
  • Proteolysis
  • RNA Interference
  • Receptors, Cell Surface
  • Sequence Homology, Amino Acid
  • Tetradecanoylphorbol Acetate
  • Tumor Necrosis Factor-alpha


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