title: The complement inhibitor, CRIT, undergoes clathrin-dependent endocytosis.
abstract: Complement C2 receptor inhibitor trispanning (CRIT) is a receptor for the second component of complement and is found in various tissues and hemopoietic cells. On binding to CRIT, C2 cannot be activated to potentially form a variant-C3 convertase as it is rendered non-cleavable by C1s. CRIT thus limits the amount of C3 convertase formed on the cell surface. In this study we have shown, using flow cytometry and immunofluorescence microscopy, that human CRIT undergoes endocytosis from the plasma membrane. The endocytosis, possibly ligand mediated, occurs via clathrin-coated pits as it can be inhibited by prior incubation of cells in hypertonic medium or with chlorpromazine, at 37 degrees C. However, inhibition of endocytosis was not possible after treatment with nystatin, or filipin, inhibitors of caveolae/raft-dependent endocytosis. In the presence of C2 alone, CRIT associates with the adapter protein, beta-arrestin-2, and whether in association with C2 or not, then appears in the perinuclear region, but does not appear to be translocated into the nucleus. Apart from the C3aR and C5aR that internalize the anaphylatoxic peptides, this is the first report of the internalization via the clathrin pathway of a receptor for a complement serum protein.
authors.affiliation: University Hospital Basel, Immunonephrology, Department of Research, 4031 Basel, Switzerland. j.inal@londonmet.ac.uk
authors.forename: Jameel
authors.initials: J
authors.lastname: Inal
authors.forename: Sylvie
authors.initials: S
authors.lastname: Miot
authors.forename: Jürg A
authors.initials: JA
authors.lastname: Schifferli
mesh_terms: D019390:Arrestins / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism* / Q000494:pharmacology; D022163:Clathrin-Coated Vesicles / Q000737:chemistry / Q000502:physiology*; D003078:Colchicine / Q000494:pharmacology; D003175:Complement C2 / Q000494:pharmacology; D003169:Complement Inactivator Proteins / Q000494:pharmacology / Q000502:physiology*; D004705:Endocytosis / Q000502:physiology*; D005372:Filipin / Q000494:pharmacology; D006801:Humans; D019169:Jurkat Cells; D008024:Ligands; D009761:Nystatin / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction / Q000502:physiology*; D014443:Tyrosine / Q000737:chemistry
chemical_list: D019390:Arrestins; C505022:CRIT protein, human; D002352:Carrier Proteins; D003175:Complement C2; D003169:Complement Inactivator Proteins; D008024:Ligands; D010750:Phosphoproteins; D009761:Nystatin; D014443:Tyrosine; D005372:Filipin; D003078:Colchicine