Biochemical Characterization of the Murine S100A9 (MRP14) Protein Suggests That It Is Functionally Equivalent to Its Human Counterpart Despite Its Low Degree of Sequence Homology

Eur J Biochem. 2000 Jan;267(2):560-5. doi: 10.1046/j.1432-1327.2000.01040.x.

Abstract

Due to the low degree of sequence similarity it has been speculated that murine and human S100A9 (MRP14), an inflammatory marker protein belonging to the S100 protein family, may have different cellular functions in mouse and man. The present study was undertaken to investigate the murine S100A9 protein (mS100A9) biochemically. We demonstrate that in murine peripheral CD11b+ cells up to 20% of the protein of the cytosolic fraction consists of mS100A9 and that several minor mS100A9 isoforms are present. Cell fractionation experiments with CD11b+ murine leukocytes showed that mS100A9 is found in the cytosol as well as in the insoluble fraction. Transient expression of a green fluorescence protein-mS100A9 fusion in mammalian cells revealed that mS100A9 is localized in neither the nucleus nor the vesicles. Recombinantly expressed murine S100A9 interacts in vitro with murine and human S100A8 in an in vitro glutathione S-transferase pull-down assay. Homodimerization was not observed. For further biochemical analysis the myeloid 32D cell line is presented as a suitable model, to study murine myeloid expressed S100 proteins. Both murine S100A9 and its dimerization partner mS100A8 are expressed at the onset of granulocyte-colony stimulating factor induced myeloid differentiation. Substantial amounts of this complex are constitutively secreted by granulocytic 32D cells into the medium. In summary, these data suggest, that the human and murine S100A9 may share a higher degree of functional homology than of sequence similarity.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Differentiation / drug effects
  • Antigens, Differentiation / genetics*
  • Antigens, Differentiation / metabolism*
  • Blotting, Western
  • Calcium-Binding Proteins / metabolism
  • Calgranulin A
  • Calgranulin B
  • Cell Line
  • Cytosol / metabolism
  • Granulocyte Colony-Stimulating Factor / pharmacology
  • Granulocytes / metabolism*
  • Humans
  • L-Lactate Dehydrogenase / metabolism
  • Leukocytes / metabolism
  • Mice
  • Precipitin Tests
  • Protein Isoforms / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • S100 Proteins / drug effects
  • S100 Proteins / genetics*
  • S100 Proteins / metabolism*
  • Sequence Homology, Amino Acid
  • Subcellular Fractions

Substances

  • Antigens, Differentiation
  • Calcium-Binding Proteins
  • Calgranulin A
  • Calgranulin B
  • Protein Isoforms
  • Recombinant Fusion Proteins
  • S100 Proteins
  • Granulocyte Colony-Stimulating Factor
  • L-Lactate Dehydrogenase