Redox regulation of CD21 shedding involves signaling via PKC and indicates the formation of a juxtamembrane stalk

J Cell Sci. 2006 Jul 15;119(Pt 14):2892-902. doi: 10.1242/jcs.02984. Epub 2006 Jun 27.

Abstract

Soluble CD21 (sCD21), released from the plasma membrane by proteolytic cleavage (shedding) of its extracellular domain (ectodomain) blocks B cell/follicular dendritic cell interaction and activates monocytes. We show here that both serine- and metalloproteases are involved in CD21 shedding. Using the oxidant pervanadate to mimic B cell receptor activation and thiol antioxidants such as N-acetylcysteine (NAC) and glutathione (GSH) we show that CD21 shedding is a redox-regulated process inducible by oxidation presumably through activation of a tyrosine kinase-mediated signal pathway involving protein kinase C (PKC), and by reducing agents that either directly activate the metalloprotease and/or modify intramolecular disulfide bridges within CD21 and thereby facilitate access to the cleavage site. Lack of short consensus repeat 16 (SCR16) abolishes CD21 shedding, and opening of the disulfide bridge between cys-2 (Cys941) and cys-4 (Cys968) of SCR16 is a prerequisite for CD21 shedding. Replacing these cysteines with selenocysteines (thereby changing the redox potential from -180 to -381 mV) results in a loss of inducible CD21 shedding, and removing this bridge by exchanging these cysteines with methionines increases CD21 shedding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antioxidants / pharmacology
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Cysteine / metabolism
  • Disulfides / metabolism
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Humans
  • Ionomycin / pharmacology
  • Metalloproteases / metabolism
  • Models, Biological
  • Molecular Sequence Data
  • Oxidation-Reduction / drug effects
  • Protein Kinase C / metabolism*
  • Protein Processing, Post-Translational* / drug effects
  • Receptors, Complement 3d / chemistry
  • Receptors, Complement 3d / immunology*
  • Serine Endopeptidases / metabolism
  • Signal Transduction*
  • Sulfhydryl Compounds / metabolism
  • Tetradecanoylphorbol Acetate / pharmacology
  • Vanadates / pharmacology

Substances

  • Antioxidants
  • Disulfides
  • Receptors, Complement 3d
  • Sulfhydryl Compounds
  • pervanadate
  • Vanadates
  • Ionomycin
  • Protein Kinase C
  • Metalloproteases
  • Serine Endopeptidases
  • Cysteine
  • Tetradecanoylphorbol Acetate