[en] Collagens are the main structural component of the extracellular matrix and provide biomechanical properties to connective tissues. A critical step in collagen fibril formation is the proteolytic removal of N- and C-terminal propeptides from procollagens by metalloproteinases of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) and BMP1 (bone morphogenetic protein 1)/Tolloid-like families, respectively. BMP1 also cleaves and activates the lysyl oxidase (LOX) precursor, the enzyme catalyzing the initial step in the formation of covalent collagen cross-links, an essential process for fibril stabilization. In this study, using murine skin fibroblasts and HEK293 cells, along with immunoprecipitation, LOX enzymatic activity, solid-phase binding assays, and proteomics analyses, we report that the LOX precursor is proteolytically processed by the procollagen N-proteinases ADAMTS2 and ADAMTS14 between Asp-218 and Tyr-219, 50 amino acids downstream of the BMP1 cleavage site. We noted that the LOX sequence between the BMP1- and ADAMTS-processing sites contains several conserved tyrosine residues, of which some are post-translationally modified by tyrosine O-sulfation and contribute to binding to collagen. Taken together, these findings unravel an additional level of regulation in the formation of collagen fibrils. They point to a mechanism that controls the binding of LOX to collagen and is based on differential BMP1- and ADAMTS2/14-mediated cleavage of a tyrosine-sulfated domain.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Rosell-García, Tamara
Paradela, Alberto
Bravo, Gema
Dupont, Laura ; Université de Liège - ULiège > GIGA Cancer - Connective Tissue Biology
Engel, J., and Chiquet, M. (2011) in The Extracellular Matrix: an Overview (Mecham, R. P., ed) pp. 1-39, Springer-Verlag, Berlin, Heidelberg
Kadler, K. E., Baldock, C., Bella, J., and Boot-Handford, R. P. (2007) Collagens at a glance. J. Cell Sci. 120, 1955-1958
Canty, E. G., and Kadler, K. E. (2005) Procollagen trafficking, processing and fibrillogenesis. J. Cell Sci. 118, 1341-1353
Eyre, D. R., and Wu, J. J. (2005) Collagen cross-links. Top. Curr. Chem. 247, 207-229
Grau-Bové, X., Ruiz-Trillo, I., and Rodriguez-Pascual, F. (2015) Origin and evolution of lysyl oxidases. Sci. Rep. 5, 10568
Rodriguez-Pascual, F., and Slatter, D. A. (2016) Collagen cross-linking: insights on the evolution of metazoan extracellular matrix. Sci. Rep. 6, 37374
Trackman, P. C., Bedell-Hogan, D., Tang, J., and Kagan, H. M. (1992) Post-translational glycosylation and proteolytic processing of a lysyl oxidase precursor. J. Biol. Chem. 267, 8666-8671
Uzel, M. I., Scott, I. C., Babakhanlou-Chase, H., Palamakumbura, A. H., Pappano, W. N., Hong, H. H., Greenspan, D. S., and Trackman, P. C. (2001) Multiple bone morphogenetic protein 1-related mammalian metalloproteinases process pro-lysyl oxidase at the correct physiological site and control lysyl oxidase activation in mouse embryo fibroblast cultures. J. Biol. Chem. 276, 22537-22543
Cronshaw, A. D., Fothergill-Gilmore, L. A., and Hulmes, D. J. (1995) The proteolytic processing site of the precursor of lysyl oxidase. Biochem. J. 306, 279-284
Panchenko, M. V., Stetler-Stevenson, W. G., Trubetskoy, O. V., Gacheru, S. N., and Kagan, H. M. (1996) Metalloproteinase activity secreted by fibrogenic cells in the processing of prolysyl oxidase: potential role of procollagen c-proteinase. J. Biol. Chem. 271, 7113-7119
Kagan, H. M., Sullivan, K. A., Olsson, T. A., 3rd., and Cronlund, A. L. (1979) Purification and properties of four species of lysyl oxidase from bovine aorta. Biochem. J. 177, 203-214
Vidal, G. P., Shieh, J. J., and Yasunobu, K. T. (1975) Immunological studies of bovine aorta lysyl oxidase: evidence for two forms of the enzyme. Biochem. Biophys. Res. Commun. 64, 989-995
Kuivaniemi, H. (1985) Partial characterization of lysyl oxidase from several human tissues. Biochem. J. 230, 639-643
Rosini, S., Pugh, N., Bonna, A. M., Hulmes, D. J. S., Farndale, R. W., and Adams, J. C. (2018) Thrombospondin-1 promotes matrix homeostasis by interacting with collagen and lysyl oxidase precursors and collagen crosslinking sites. Sci. Signal. 11, eaar2566
Bekhouche, M., Leduc, C., Dupont, L., Janssen, L., Delolme, F., Vadon-Le Goff, S., Smargiasso, N., Baiwir, D., Mazzucchelli, G., Zanella-Cleon, I., Dubail, J., De Pauw, E., Nusgens, B., Hulmes, D. J., Moali, C., and Colige, A. (2016) Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF- signaling as primary targets. FASEB J. 30, 1741-1756
Boak, A. M., Roy, R., Berk, J., Taylor, L., Polgar, P., Goldstein, R. H., and Kagan, H. M. (1994) Regulation of lysyl oxidase expression in lung fibroblasts by transforming growth factor-1 and prostaglandin E2. Am. J. Respir. Cell Mol. Biol. 11, 751-755
Dupont, L., Ehx, G., Chantry, M., Monseur, C., Leduc, C., Janssen, L., Cataldo, D., Thiry, M., Jerome, C., Thomassin, J. M., Nusgens, B., Dubail, J., Baron, F., and Colige, A. (2018) Spontaneous atopic dermatitis due to immune dysregulation in mice lacking Adamts2 and 14. Matrix Biol. 70, 140-157
Kehoe, J. W., and Bertozzi, C. R. (2000) Tyrosine sulfation: a modulator of extracellular protein-protein interactions. Chem. Biol. 7, R57-R61
Yang, Y. S., Wang, C. C., Chen, B. H., Hou, Y. H., Hung, K. S., and Mao, Y. C. (2015) Tyrosine sulfation as a protein post-translational modification. Molecules 20, 2138-2164
Atsawasuwan, P., Mochida, Y., Katafuchi, M., Tokutomi, K., Mocanu, V., Parker, C. E., and Yamauchi, M. (2011) A novel proteolytic processing of prolysyl oxidase. Connect. Tissue Res. 52, 479-486
Monigatti, F., Gasteiger, E., Bairoch, A., and Jung, E. (2002) The sulfinator: predicting tyrosine sulfation sites in protein sequences. Bioinformatics 18, 769-770
Palamakumbura, A. H., and Trackman, P. C. (2002) A fluorometric assay for detection of lysyl oxidase enzyme activity in biological samples. Anal. Biochem. 300, 245-251
Trackman, P. C., and Kagan, H. M. (1979) Nonpeptidyl amine inhibitors are substrates of lysyl oxidase. J. Biol. Chem. 254, 7831-7836
Kagan, H. M., Williams, M. A., Williamson, P. R., and Anderson, J. M. (1984) Influence of sequence and charge on the specificity of lysyl oxidase toward protein and synthetic peptide substrates. J. Biol. Chem. 259, 11203-11207
Rodriguez-Pascual, F., and Rosell-Garcia, T. (2018) Lysyl oxidases: functions and disorders. J. Glaucoma 27, S15-S19
Blouin, S., Paschalis, E., Gupta, H., Fratzl-Zelman, N., Pischon, N., Trackman, P. C., Mäki, J. M., Roschger, P., and Klaushofer, K. (2010) Bone material properties in lysyl oxidase knock-out mice. Bone 47, S80
Mäki, J. M., Räsänen, J., Tikkanen, H., Sormunen, R., Mäkikallio, K., Kivirikko, K. I., and Soininen, R. (2002) Inactivation of the lysyl oxidase gene Lox leads to aortic aneurysms, cardiovascular dysfunction, and perinatal death in mice. Circulation 106, 2503-2509
Mäki, J. M., Sormunen, R., Lippo, S., Kaarteenaho-Wiik, R., Soininen, R., and Myllyharju, J. (2005) Lysyl oxidase is essential for normal development and function of the respiratory system and for the integrity of elastic and collagen fibers in various tissues. Am. J. Pathol. 167, 927-936
Uzel, M. I., Shih, S. D., Gross, H., Kessler, E., Gerstenfeld, L. C., and Trackman, P. C. (2000) Molecular events that contribute to lysyl oxidase enzyme activity and insoluble collagen accumulation in osteosarcoma cell clones. J. Bone Miner. Res. 15, 1189-1197
Cronlund, A. L., Smith, B. D., and Kagan, H. M. (1985) Binding of lysyl oxidase to fibrils of type I collagen. Connect. Tissue Res. 14, 109-119
Siegel, R. C. (1974) Biosynthesis of collagen crosslinks: increased activity of purified lysyl oxidase with reconstituted collagen fibrils. Proc. Natl. Acad. Sci. U.S.A. 71, 4826-4830
Kanan, Y., Siefert, J. C., Kinter, M., and Al-Ubaidi, M. R. (2014) Complement factor H, vitronectin, and opticin are tyrosine-sulfated proteins of the retinal pigment epithelium. PLoS ONE 9, e105409
Onnerfjord, P., Heathfield, T. F., and Heinegård, D. (2004) Identification of tyrosine sulfation in extracellular leucine-rich repeat proteins using mass spectrometry. J. Biol. Chem. 279, 26-33
Tillgren, V., Mörgelin, M.,Ö nnerfjord, P., Kalamajski, S., and Aspberg, A. (2016) The tyrosine sulfate domain of fibromodulin binds collagen and enhances fibril formation. J. Biol. Chem. 291, 23744-23755
Kalamajski, S., Bihan, D., Bonna, A., Rubin, K., and Farndale, R. W. (2016) Fibromodulin interacts with collagen cross-linking sites and activates lysyl oxidase. J. Biol. Chem. 291, 7951-7960
Svensson, L., Aszódi, A., Reinholt, F. P., Fässler, R., Heinegård, D., and Oldberg, A. (1999) Fibromodulin-null mice have abnormal collagen fibrils, tissue organization, and altered lumican deposition in tendon. J. Biol. Chem. 274, 9636-9647
Kalamajski, S., Liu, C., Tillgren, V., Rubin, K., Oldberg, Å., Rai, J., Weis, M., and Eyre, D. R. (2014) Increased C-telopeptide cross-linking of tendon type I collagen in fibromodulin-deficient mice. J. Biol. Chem. 289, 18873-18879
Moali, C., and Hulmes, D. J. (2012) in Extracellular Matrix: Pathobiology and Signaling (Karamanos, N., ed) pp. 539-561, Walter de Gruyter, Berlin/Boston
Rawlings, N. D., Waller, M., Barrett, A. J., and Bateman, A. (2014) MEROPS: the database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Res. 42, D503-D509
Marini, J. C., Forlino, A., Bächinger, H. P., Bishop, N. J., Byers, P. H., Paepe, A., Fassier, F., Fratzl-Zelman, N., Kozloff, K. M., Krakow, D., Montpetit, K., and Semler, O. (2017) Osteogenesis imperfecta. Nat. Rev. Dis. Primers 3, 17052
Colige, A., Sieron, A. L., Li, S. W., Schwarze, U., Petty, E., Wertelecki, W., Wilcox, W., Krakow, D., Cohn, D. H., Reardon, W., Byers, P. H., Lapiere, C. M., Prockop, D. J., and Nusgens, B. V. (1999) Human Ehlers-Danlos syndrome type VIICand bovine dermatosparaxis are caused by mutations in the procollagen I N-proteinase gene. Am. J. Hum. Genet. 65, 308-317
Bekhouche, M., and Colige, A. (2015) The procollagen N-proteinases ADAMTS2, 3 and 14 in pathophysiology. Matrix Biol. 44-46, 46-53
Le Goff, C., Somerville, R. P., Kesteloot, F., Powell, K., Birk, D. E., Colige, A. C., and Apte, S. S. (2006) Regulation of procollagen amino-propeptide processing during mouse embryogenesis by specialization of homologous ADAMTS proteases: insights on collagen biosynthesis and dermatosparaxis. Development 133, 1587-1596
Guo, D.-C., Regalado, E. S., Gong, L., Duan, X., Santos-Cortez, R. L. P., Arnaud, P., Ren, Z., Cai, B., Hostetler, E. M., Moran, R., Liang, D., Estrera, A., Safi, H. J., University of Washington Center for Mendelian Genomics, Leal, S. M., et al. (2016) LOX mutations predispose to thoracic aortic aneurysms and dissections. Circ. Res. 118, 928-934
Lee, V. S., Halabi, C. M., Hoffman, E. P., Carmichael, N., Leshchiner, I., Lian, C. G., Bierhals, A. J., Vuzman, D., Brigham Genomic Medicine, Mecham, R. P., Frank, N. Y., and Stitziel, N. O. (2016) Loss of function mutation in LOX causes thoracic aortic aneurysm and dissection in humans. Proc. Natl. Acad. Sci. U.S.A. 113, 8759-8764
Hunt, S. E., McLaren, W., Gil, L., Thormann, A., Schuilenburg, H., Sheppard, D., Parton, A., Armean, I. M., Trevanion, S. J., Flicek, P., and Cunningham, F. (2018) Ensembl variation resources. Database 2018
Busnadiego, O., González-Santamaría, J., Lagares, D., Guinea-Viniegra, J., Pichol-Thievend, C., Muller, L., and Rodríguez-Pascual, F. (2013) LOXL4 is induced by transforming growth factor 1 through Smad and JunB/Fra2 and contributes to vascular matrix remodeling. Mol. Cell. Biol. 33, 2388-2401
Puig, M., Lugo, R., Gabasa, M., Giménez, A., Velásquez, A., Galgoczy, R., Ramírez, J., Gómez-Caro, A., Busnadiego, Ó., Rodríguez-Pascual, F., Gascón, P., Reguart, N., and Alcaraz, J. (2015) Matrix stiffening and 1 integrin drive subtype-specific fibroblast accumulation in lung cancer. Mol. Cancer Res. 13, 161-173
Rodríguez-Pascual, F., Redondo-Horcajo, M., and Lamas, S. (2003) Functional cooperation between Smad proteins and activator protein-1 regulates transforming growth factor-mediated induction of endothelin-1 expression. Circ. Res. 92, 1288-1295
Martínez-Glez, V., Valencia, M., Caparrós-Martín, J. A., Aglan, M., Temtamy, S., Tenorio, J., Pulido, V., Lindert, U., Rohrbach, M., Eyre, D., Giunta, C., Lapunzina, P., and Ruiz-Perez, V. L. (2012) Identification of a mutation causing deficient BMP1/mTLD proteolytic activity in autosomal recessive osteogenesis imperfecta. Hum. Mutat. 33, 343-350
Colige, A., Ruggiero, F., Vandenberghe, I., Dubail, J., Kesteloot, F., Van Beeumen, J., Beschin, A., Brys, L., Lapiere, C. M., and Nusgens, B. (2005) Domains and maturation processes that regulate the activity of ADAMTS-2, a metalloproteinase cleaving the aminopropeptide of fibrillar procollagens types I-III and V. J. Biol. Chem. 280, 34397-34408
Wessel, D., and Flügge, U. I. (1984) Amethod for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal. Biochem. 138, 141-143
Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 68, 850-858
Zhang, J., Xin, L., Shan, B., Chen, W., Xie, M., Yuen, D., Zhang, W., Zhang, Z., Lajoie, G. A., and Ma, B. (2012) PEAKS DB: de novo sequencing assisted database search for sensitive and accurate peptide identification. Mol. Cell. Proteomics 11, M111.010587
Mould, P. A. (2009) Solid phase assays for studying ECM protein-protein interactions. Methods Mol. Biol. 522, 195-200