title: Internalization of exogenous cystatin F supresses cysteine proteases and induces the accumulation of single-chain cathepsin L by multiple mechanisms.
abstract: Cystatin F is an unusual member of the cystatin family of protease inhibitors, which is made as an inactive dimer and becomes activated by proteolysis in the endo/lysosome pathway of the immune cells that produce it. However a proportion is secreted and can be taken up and activated by other cells. We show here that cystatin F acquired in this way induces a dramatic accumulation of the single-chain form of cathepsin L (CatL). Cystatin F was observed in the same cellular compartments as CatL and was tightly complexed with CatL as determined by co-precipitation studies. The observed accumulation of single-chain CatL was partly due to cystatin F-mediated inhibition of the putative single-chain to two-chain CatL convertase AEP/legumain and partly to general suppression of cathepsin activity. Thus, cystatin F stabilizes CatL leading to the dramatic accumulation of an inactive complex composed either of the single-chain or two-chain form depending on the capacity of cystatin F to inhibit AEP. Cross-transfer of cystatin F from one cell to another may therefore attenuate potentially harmful effects of excessive CatL activity while paradoxically, inducing accumulation of CatL protein. Finally, we confirmed earlier data (Beers, C., Honey, K., Fink, S., Forbush, K., and Rudensky, A. (2003) J. Exp. Med. 197, 169-179) showing a loss of CatL activity, but not of CatL protein, in macrophages activated with IFNγ. However, we found equivalent loss of CatL activity in wild type and cystatin F-null macrophages suggesting that an inhibitory activity other than cystatin F quenches CatL activity in activated macrophages.
authors.affiliation: Division of Cell Signaling and Immunology, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, United Kingdom.
authors.forename: Jeff D
authors.initials: JD
authors.lastname: Colbert
authors.affiliation: Division of Cell Signaling and Immunology, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, United Kingdom.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Matthews
authors.affiliation: Faculty of Pharmacy, University of Ljubljana, Askerceva 7, 1000 Ljubljana, Slovenia.
authors.forename: Janko
authors.initials: J
authors.lastname: Kos
authors.affiliation: Division of Cell Signaling and Immunology, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, United Kingdom. Electronic address: c.watts@dundee.ac.uk.
authors.forename: Colin
authors.initials: C
authors.lastname: Watts
mesh_terms: D000818:Animals; D001216:Asparagine / Q000737:chemistry; D001854:Bone Marrow Cells / Q000166:cytology; D056668:Cathepsin L / Q000378:metabolism*; D055316:Cystatin C / Q000378:metabolism; D015891:Cystatins / Q000378:metabolism*; D003713:Dendritic Cells / Q000166:cytology*; D004705:Endocytosis; D010450:Endopeptidases / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D007700:Kinetics; D008247:Lysosomes / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D010447:Peptide Hydrolases / Q000378:metabolism; D013154:Spleen / Q000166:cytology
chemical_list: D055316:Cystatin C; D015891:Cystatins; C550521:cystatin F, mouse; D001216:Asparagine; D010450:Endopeptidases; D010447:Peptide Hydrolases; D056668:Cathepsin L