title: Cysteine Cathepsins Activate ELR Chemokines and Inactivate Non-ELR Chemokines.
abstract: Cysteine cathepsins are primarily lysosomal proteases involved in general protein turnover, but they also have specific proteolytic functions in antigen presentation and bone remodeling. Cathepsins are most stable at acidic pH, although growing evidence indicates that they have physiologically relevant activity also at neutral pH. Post-translational proteolytic processing of mature chemokines is a key, yet underappreciated, level of chemokine regulation. Although the role of selected serine proteases and matrix metalloproteases in chemokine processing has long been known, little has been reported about the role of cysteine cathepsins. Here we evaluated cleavage of CXC ELR (CXCL1, -2, -3, -5, and -8) and non-ELR (CXCL9-12) chemokines by cysteine cathepsins B, K, L, and S at neutral pH by high resolution Tris-Tricine SDS-PAGE and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Whereas cathepsin B cleaved chemokines especially in the C-terminal region, cathepsins K, L, and S cleaved chemokines at the N terminus with glycosaminoglycans modulating cathepsin processing of chemokines. The functional consequences of the cleavages were determined by Ca(2+) mobilization and chemotaxis assays. We show that cysteine cathepsins inactivate and in some cases degrade non-ELR CXC chemokines CXCL9-12. In contrast, cathepsins specifically process ELR CXC chemokines CXCL1, -2, -3, -5, and -8 N-terminally to the ELR motif, thereby generating agonist forms. This study suggests that cysteine cathepsins regulate chemokine activity and thereby leukocyte recruitment during protective or pathological inflammation.
authors.affiliation: From the Department of Biochemistry and Molecular and Structural Biology, Jozef Stefan Institute, SI-1000 Ljubljana, Slovenia.
authors.forename: Urska
authors.initials: U
authors.lastname: Repnik
authors.affiliation: the Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Amanda E
authors.initials: AE
authors.lastname: Starr
authors.affiliation: the Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada, the Department of Oral Biological and Medical Sciences, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada, chris.overall@ubc.ca.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Overall
authors.affiliation: From the Department of Biochemistry and Molecular and Structural Biology, Jozef Stefan Institute, SI-1000 Ljubljana, Slovenia, the Centre of Excellence for Integrated Approaches in Chemistry and Biology of Proteins, SI-1000 Ljubljana, Slovenia, and the Faculty of Chemistry and Chemical Technology, University of Ljubljana, SI-1000 Ljubljana, Slovenia boris.turk@ijs.si.
authors.forename: Boris
authors.initials: B
authors.lastname: Turk
mesh_terms: D002118:Calcium / Q000378:metabolism; D002403:Cathepsins / Q000378:metabolism*; D002460:Cell Line; D002465:Cell Movement; D018925:Chemokines / Q000378:metabolism*; D002633:Chemotaxis; D003545:Cysteine / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D006025:Glycosaminoglycans / Q000378:metabolism; D006098:Granulocytes / Q000166:cytology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007249:Inflammation; D013058:Mass Spectrometry; D009504:Neutrophils / Q000166:cytology; D017434:Protein Structure, Tertiary; D023063:Receptors, Interleukin-8B / Q000378:metabolism; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D018925:Chemokines; D006025:Glycosaminoglycans; D023063:Receptors, Interleukin-8B; D002403:Cathepsins; D003545:Cysteine; D002118:Calcium
keywords: CXC chemokine; chemokine; chemokine processing; cysteine cathepsin; glycosaminoglycan; inflammation; mass spectrometry (MS); proteolysis
references.citation: Biosci Rep. 2006 Oct;26(5):325-39
references.pmid: 17024562
references.citation: Cell Cycle. 2007 Jan 1;6(1):60-4
references.pmid: 17245112
references.citation: Eur J Biochem. 1999 Mar;260(2):421-9
references.pmid: 10095777
references.citation: J Biol Chem. 1999 Oct 22;274(43):30433-8
references.pmid: 10521421
references.citation: FEBS Lett. 1999 Oct 15;459(3):358-62
references.pmid: 10526165
references.citation: Biochem Soc Trans. 2006 Jun;34(Pt 3):422-6
references.pmid: 16709177
references.citation: Curr Pharm Des. 2007;13(4):387-403
references.pmid: 17311556
references.citation: Nat Rev Mol Cell Biol. 2007 Mar;8(3):245-57
references.pmid: 17299501
references.citation: PLoS One. 2007;2(3):e312
references.pmid: 17375198
references.citation: Mol Cell Proteomics. 2007 Apr;6(4):611-23
references.pmid: 17200105
references.citation: Blood. 2007 Jul 1;110(1):37-44
references.pmid: 17363734
references.citation: J Biol Chem. 2007 Nov 9;282(45):33076-85
references.pmid: 17726009
references.citation: Biochimie. 2008 Feb;90(2):380-6
references.pmid: 17991442
references.citation: Pharmacol Ther. 2008 Nov;120(2):197-217
references.pmid: 18793669
references.citation: J Neurosci. 2009 May 27;29(21):6945-54
references.pmid: 19474321
references.citation: Methods Enzymol. 2009;461:281-307
references.pmid: 19480924
references.citation: J Biol Chem. 2009 Aug 14;284(33):21783-7
references.pmid: 19473965
references.citation: Cell Physiol Biochem. 2010;25(2-3):315-24
references.pmid: 20110692
references.citation: Genes Dev. 2010 Feb 1;24(3):241-55
references.pmid: 20080943
references.citation: Bioorg Med Chem. 2011 Feb 1;19(3):1055-61
references.pmid: 21130662
references.citation: Exp Cell Res. 2011 Mar 10;317(5):642-54
references.pmid: 21146523
references.citation: Exp Cell Res. 2011 Mar 10;317(5):590-601
references.pmid: 21223963
references.citation: J Proteome Res. 2011 Dec 2;10(12):5363-73
references.pmid: 21967108
references.citation: Biochim Biophys Acta. 2012 Jan;1824(1):68-88
references.pmid: 22024571
references.citation: J Biol Chem. 2012 Feb 17;287(8):5848-60
references.pmid: 22147696
references.citation: EMBO J. 2012 Apr 4;31(7):1630-43
references.pmid: 22367392
references.citation: J Biol Chem. 2012 Apr 13;287(16):13382-95
references.pmid: 22367194
references.citation: Arthritis Rheum. 2012 Jun;64(6):2038-47
references.pmid: 22213084
references.citation: Stem Cells Dev. 2012 Jul 20;21(11):1924-35
references.pmid: 22066471
references.citation: Biol Chem. 2013 Feb;394(2):307-16
references.pmid: 23152404
references.citation: Arthritis Rheum. 2013 Nov;65(11):2814-25
references.pmid: 23897050
references.citation: PLoS Biol. 2014 May;12(5):e1001869
references.pmid: 24865846
references.citation: Biochim Biophys Acta. 2014 Aug;1840(8):2560-70
references.pmid: 24680817
references.citation: Cell Rep. 2014 Oct 23;9(2):618-32
references.pmid: 25310974
references.citation: J Immunol. 2000 Apr 1;164(7):3774-82
references.pmid: 10725737
references.citation: J Leukoc Biol. 2000 Jul;68(1):1-8
references.pmid: 10914483
references.citation: Science. 2000 Aug 18;289(5482):1202-6
references.pmid: 10947989
references.citation: Blood. 2000 Oct 15;96(8):2673-81
references.pmid: 11023497
references.citation: J Biol Chem. 2001 Jan 12;276(2):944-51
references.pmid: 11016923
references.citation: J Biol Chem. 2001 Nov 23;276(47):43503-8
references.pmid: 11571304
references.citation: Blood. 2001 Dec 15;98(13):3554-61
references.pmid: 11739156
references.citation: Arthritis Rheum. 2002 Mar;46(3):663-74
references.pmid: 11920402
references.citation: J Leukoc Biol. 2002 Jul;72(1):183-91
references.pmid: 12101279
references.citation: Blood. 2002 Aug 15;100(4):1160-7
references.pmid: 12149192
references.citation: J Biol Chem. 2002 Aug 9;277(32):28669-76
references.pmid: 12039963
references.citation: Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1885-90
references.pmid: 12571364
references.citation: Biochim Biophys Acta. 2003 Jun 26;1649(1):30-9
references.pmid: 12818188
references.citation: Nat Neurosci. 2003 Oct;6(10):1064-71
references.pmid: 14502291
references.citation: Methods. 2004 Feb;32(2):199-206
references.pmid: 14698633
references.citation: J Biol Chem. 2004 Feb 13;279(7):5470-9
references.pmid: 14645229
references.citation: J Exp Med. 1989 Nov 1;170(5):1745-50
references.pmid: 2681518
references.citation: Biochem J. 1989 Dec 1;264(2):467-73
references.pmid: 2690828
references.citation: Biochem Biophys Res Commun. 1995 Dec 14;217(2):624-31
references.pmid: 7503744
references.citation: Biochemistry. 1999 Jan 12;38(2):636-42
references.pmid: 9888803
references.citation: Eur J Biochem. 1999 Feb;259(3):926-32
references.pmid: 10092883
references.citation: J Immunol. 2006 Jun 1;176(11):6512-22
references.pmid: 16709808
references.citation: FEBS Lett. 2008 Mar 5;582(5):819-25
references.pmid: 18275857
references.citation: J Biol Chem. 2008 May 23;283(21):14453-60
references.pmid: 18362148
references.citation: J Biol Chem. 2008 Jul 11;283(28):19389-99
references.pmid: 18411283
references.citation: Cancer Lett. 2008 Aug 28;267(2):226-44
references.pmid: 18579287
references.citation: Blood. 2008 Oct 15;112(8):3455-64
references.pmid: 18660381
references.citation: FEBS Lett. 2005 Feb 14;579(5):1285-90
references.pmid: 15710427
references.citation: J Biol Chem. 1996 Aug 23;271(34):20545-50
references.pmid: 8702798
references.citation: J Biol Chem. 1997 Jan 10;272(2):1197-202
references.pmid: 8995421
references.citation: J Exp Med. 1996 Sep 1;184(3):963-9
references.pmid: 9064356
references.citation: Br J Rheumatol. 1997 Jul;36(7):735-43
references.pmid: 9255106
references.citation: Methods Enzymol. 1997;287:233-50
references.pmid: 9330326
references.citation: Biol Chem. 1998 Feb;379(2):137-47
references.pmid: 9524065
references.citation: Nat Rev Cancer. 2006 Oct;6(10):764-75
references.pmid: 16990854