Modulation of mannose receptor activity by proteolysis.
Macrophages express a receptor on the cell surface that functions to clear glycoproteins from the extracellular milieu. The activity of this receptor is sensitive to treatment with trypsin. In inflammatory situations, macrophages are activated and exposed to increased levels of extracellular proteases. Under these conditions, mannose receptor activity on the macrophages is diminished. We therefore decided to study the effects of trypsin treatment on the structure and activity of cell-associated and purified receptor that might contribute to the activation-associated receptor down-regulation. Trypsin treatment (1 microgram/ml for 3 h) resulted in the production of a 140 kDa, trypsin-resistant fragment from both intact cells and isolated receptor. This fragment was no longer able to bind ligand. The remaining 35 kDa fragment apparently is further degraded into smaller fragments, since no evidence of this domain was found on Coomassie Blue-stained gels. The 140 kDa fragment retained immunoreactivity and contained at least a portion of the iodinated tyrosine residues following surface labelling with Na125I. Neither calcium nor ligand protected the receptor from proteolysis. In addition, prior treatment with oxidants did not increase the susceptibility of the receptor to trypsin digestion. We conclude from these results that the macrophage mannose receptor is clipped by the serine protease trypsin at the cell surface, resulting in the release and further degradation of the binding domain, and the production of a membrane-associated 140 kDa fragment. This trypsin-mediated down-regulation of receptor activity might be important in controlling glycoprotein clearance during inflammation.
Shepherd
V L
VL
VA Medical Center, Nashville, TN 37212.
Abdolrasulnia
R
R
Stephenson
J
J
Crenshaw
C
C
D000818:Animals; D001665:Binding Sites; D001854:Bone Marrow Cells; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D037181:Lectins, C-Type; D008264:Macrophages / Q000378:metabolism; D008351:Mannans / Q000378:metabolism; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D008970:Molecular Weight; D010449:Peptide Mapping; D010529:Peritoneal Cavity / Q000166:cytology; D051381:Rats; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014357:Trypsin / Q000378:metabolism* / Q000494:pharmacology
D037181:Lectins, C-Type; D008351:Mannans; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D006861:Hydrogen Peroxide; D014357:Trypsin; D002118:Calcium