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. 2004 Feb 1;377(Pt 3):787–795. doi: 10.1042/BJ20030896

ADAMTS4 (aggrecanase-1) cleaves human brain versican V2 at Glu405-Gln406 to generate glial hyaluronate binding protein.

Jennifer Westling 1, Paul E Gottschall 1, Vivian P Thompson 1, Amber Cockburn 1, George Perides 1, Dieter R Zimmermann 1, John D Sandy 1
PMCID: PMC1223897  PMID: 14561220

Abstract

Human brain tissue from cerebellum and hippocampus was obtained between 2 h and 24 h post mortem and, after extraction in the presence of proteinase inhibitors, proteoglycans were purified by anion-exchange chromatography. The versican component was characterized by Western analysis with antibodies to the N-terminal peptide (LF99), the N-terminal globular domain (12C5) and the two GAG (glycosaminoglycan) attachment regions (anti-GAG-alpha and anti-GAG-beta). The results indicated that versican V2 is the major variant in all brain samples, and that it exists as the full-length form and also as at least six C-terminally truncated forms. The major immunoreactive species present is a 64 kDa product, which we identified by biochemical and immunological analysis as the brain protein previously termed GHAP (glial hyaluronate binding protein) [Perides, Lane, Andrews, Dahl and Bignami (1989) J. Biol. Chem. 264, 5981-5987]. Immunological analysis of purified human GHAP using a new anti-neoepitope antiserum (JSCNIV) showed that its C-terminal sequence is NIVSFE(405), and digestion of human cerebellum proteoglycans with ADAMTS4 (aggrecanase-1, where ADAMTS, a disintegrin and metalloproteinase with thrombospondin-1-like motifs) indicated that GHAP is a product of cleavage of versican V0 or V2 at the Glu(405)-Gln(406) bond. Since human cerebellum extracts contained multiple forms of ADAMTS4 protein on Western analysis, these data suggest that one or more members of the 'aggrecanase' group of the ADAMTS family (ADAMTS 1, 4, 5 and 9) are responsible for turnover of versican V2 in the adult human brain.

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Selected References

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  1. Asher R., Perides G., Vanderhaeghen J. J., Bignami A. Extracellular matrix of central nervous system white matter: demonstration of an hyaluronate-protein complex. J Neurosci Res. 1991 Mar;28(3):410–421. doi: 10.1002/jnr.490280314. [DOI] [PubMed] [Google Scholar]
  2. Asher Richard A., Morgenstern Daniel A., Shearer Morven C., Adcock Kathryn H., Pesheva Penka, Fawcett James W. Versican is upregulated in CNS injury and is a product of oligodendrocyte lineage cells. J Neurosci. 2002 Mar 15;22(6):2225–2236. doi: 10.1523/JNEUROSCI.22-06-02225.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bignami A., LeBlanc A., Perides G. A role for extracellular matrix degradation and matrix metalloproteinases in senile dementia? Acta Neuropathol. 1994;87(3):308–312. doi: 10.1007/BF00296747. [DOI] [PubMed] [Google Scholar]
  4. Delpech A., Girard N., Delpech B. Localization of hyaluronectin in the nervous system. Brain Res. 1982 Aug 12;245(2):251–257. doi: 10.1016/0006-8993(82)90807-1. [DOI] [PubMed] [Google Scholar]
  5. Dours-Zimmermann M. T., Zimmermann D. R. A novel glycosaminoglycan attachment domain identified in two alternative splice variants of human versican. J Biol Chem. 1994 Dec 30;269(52):32992–32998. [PubMed] [Google Scholar]
  6. Fawcett J. W., Asher R. A. The glial scar and central nervous system repair. Brain Res Bull. 1999 Aug;49(6):377–391. doi: 10.1016/s0361-9230(99)00072-6. [DOI] [PubMed] [Google Scholar]
  7. Flannery Carl R., Zeng Weilan, Corcoran Chris, Collins-Racie Lisa A., Chockalingam Priya S., Hebert Tracy, Mackie Stewart A., McDonagh Thomas, Crawford Tara K., Tomkinson Kathy N. Autocatalytic cleavage of ADAMTS-4 (Aggrecanase-1) reveals multiple glycosaminoglycan-binding sites. J Biol Chem. 2002 Aug 28;277(45):42775–42780. doi: 10.1074/jbc.M205309200. [DOI] [PubMed] [Google Scholar]
  8. Gao Gui, Westling Jennifer, Thompson Vivian P., Howell Troy D., Gottschall Paul E., Sandy John D. Activation of the proteolytic activity of ADAMTS4 (aggrecanase-1) by C-terminal truncation. J Biol Chem. 2002 Jan 16;277(13):11034–11041. doi: 10.1074/jbc.M107443200. [DOI] [PubMed] [Google Scholar]
  9. Herndon M. E., Lander A. D. A diverse set of developmentally regulated proteoglycans is expressed in the rat central nervous system. Neuron. 1990 Jun;4(6):949–961. doi: 10.1016/0896-6273(90)90148-9. [DOI] [PubMed] [Google Scholar]
  10. Hörber C., Büttner F. H., Kern C., Schmiedeknecht G., Bartnik E. Truncation of the amino-terminus of the recombinant aggrecan rAgg1mut leads to reduced cleavage at the aggrecanase site. Efficient aggrecanase catabolism may depend on multiple substrate interactions. Matrix Biol. 2000 Nov;19(6):533–543. doi: 10.1016/s0945-053x(00)00113-x. [DOI] [PubMed] [Google Scholar]
  11. Landolt R. M., Vaughan L., Winterhalter K. H., Zimmermann D. R. Versican is selectively expressed in embryonic tissues that act as barriers to neural crest cell migration and axon outgrowth. Development. 1995 Aug;121(8):2303–2312. doi: 10.1242/dev.121.8.2303. [DOI] [PubMed] [Google Scholar]
  12. Lemons M. L., Sandy J. D., Anderson D. K., Howland D. R. Intact aggrecan and fragments generated by both aggrecanse and metalloproteinase-like activities are present in the developing and adult rat spinal cord and their relative abundance is altered by injury. J Neurosci. 2001 Jul 1;21(13):4772–4781. doi: 10.1523/JNEUROSCI.21-13-04772.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lemons Michele L., Sandy John D., Anderson Douglas K., Howland Dena R. Intact aggrecan and chondroitin sulfate-depleted aggrecan core glycoprotein inhibit axon growth in the adult rat spinal cord. Exp Neurol. 2003 Dec;184(2):981–990. doi: 10.1016/S0014-4886(03)00383-2. [DOI] [PubMed] [Google Scholar]
  14. Matthews R. T., Gary S. C., Zerillo C., Pratta M., Solomon K., Arner E. C., Hockfield S. Brain-enriched hyaluronan binding (BEHAB)/brevican cleavage in a glioma cell line is mediated by a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) family member. J Biol Chem. 2000 Jul 28;275(30):22695–22703. doi: 10.1074/jbc.M909764199. [DOI] [PubMed] [Google Scholar]
  15. Mercuri F. A., Maciewicz R. A., Tart J., Last K., Fosang A. J. Mutations in the interglobular domain of aggrecan alter matrix metalloproteinase and aggrecanase cleavage patterns. Evidence that matrix metalloproteinase cleavage interferes with aggrecanase activity. J Biol Chem. 2000 Oct 20;275(42):33038–33045. doi: 10.1074/jbc.275.42.33038. [DOI] [PubMed] [Google Scholar]
  16. Moon L. D., Asher R. A., Rhodes K. E., Fawcett J. W. Regeneration of CNS axons back to their target following treatment of adult rat brain with chondroitinase ABC. Nat Neurosci. 2001 May;4(5):465–466. doi: 10.1038/87415. [DOI] [PubMed] [Google Scholar]
  17. Nakamura H., Fujii Y., Inoki I., Sugimoto K., Tanzawa K., Matsuki H., Miura R., Yamaguchi Y., Okada Y. Brevican is degraded by matrix metalloproteinases and aggrecanase-1 (ADAMTS4) at different sites. J Biol Chem. 2000 Dec 8;275(49):38885–38890. doi: 10.1074/jbc.M003875200. [DOI] [PubMed] [Google Scholar]
  18. Oohashi Toshitaka, Hirakawa Satoshi, Bekku Yoko, Rauch Uwe, Zimmermann Dieter R., Su Wei-Dong, Ohtsuka Aiji, Murakami Takuro, Ninomiya Yoshifumi. Bral1, a brain-specific link protein, colocalizing with the versican V2 isoform at the nodes of Ranvier in developing and adult mouse central nervous systems. Mol Cell Neurosci. 2002 Jan;19(1):43–57. doi: 10.1006/mcne.2001.1061. [DOI] [PubMed] [Google Scholar]
  19. Perides G., Asher R. A., Lark M. W., Lane W. S., Robinson R. A., Bignami A. Glial hyaluronate-binding protein: a product of metalloproteinase digestion of versican? Biochem J. 1995 Dec 1;312(Pt 2):377–384. doi: 10.1042/bj3120377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Perides G., Lane W. S., Andrews D., Dahl D., Bignami A. Isolation and partial characterization of a glial hyaluronate-binding protein. J Biol Chem. 1989 Apr 5;264(10):5981–5987. [PubMed] [Google Scholar]
  21. Perides G., Rahemtulla F., Lane W. S., Asher R. A., Bignami A. Isolation of a large aggregating proteoglycan from human brain. J Biol Chem. 1992 Nov 25;267(33):23883–23887. [PubMed] [Google Scholar]
  22. Pizzorusso Tommaso, Medini Paolo, Berardi Nicoletta, Chierzi Sabrina, Fawcett James W., Maffei Lamberto. Reactivation of ocular dominance plasticity in the adult visual cortex. Science. 2002 Nov 8;298(5596):1248–1251. doi: 10.1126/science.1072699. [DOI] [PubMed] [Google Scholar]
  23. Rodríguez-Manzaneque Juan Carlos, Westling Jennifer, Thai Shelley N-M, Luque Alfonso, Knauper Vera, Murphy Gillian, Sandy John D., Iruela-Arispe M. Luisa. ADAMTS1 cleaves aggrecan at multiple sites and is differentially inhibited by metalloproteinase inhibitors. Biochem Biophys Res Commun. 2002 Apr 26;293(1):501–508. doi: 10.1016/S0006-291X(02)00254-1. [DOI] [PubMed] [Google Scholar]
  24. Sandy J. D., Gamett D., Thompson V., Verscharen C. Chondrocyte-mediated catabolism of aggrecan: aggrecanase-dependent cleavage induced by interleukin-1 or retinoic acid can be inhibited by glucosamine. Biochem J. 1998 Oct 1;335(Pt 1):59–66. doi: 10.1042/bj3350059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sandy J. D., Verscharen C. Analysis of aggrecan in human knee cartilage and synovial fluid indicates that aggrecanase (ADAMTS) activity is responsible for the catabolic turnover and loss of whole aggrecan whereas other protease activity is required for C-terminal processing in vivo. Biochem J. 2001 Sep 15;358(Pt 3):615–626. doi: 10.1042/0264-6021:3580615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sandy J. D., Westling J., Kenagy R. D., Iruela-Arispe M. L., Verscharen C., Rodriguez-Mazaneque J. C., Zimmermann D. R., Lemire J. M., Fischer J. W., Wight T. N. Versican V1 proteolysis in human aorta in vivo occurs at the Glu441-Ala442 bond, a site that is cleaved by recombinant ADAMTS-1 and ADAMTS-4. J Biol Chem. 2001 Jan 26;276(16):13372–13378. doi: 10.1074/jbc.M009737200. [DOI] [PubMed] [Google Scholar]
  27. Schmalfeldt M., Bandtlow C. E., Dours-Zimmermann M. T., Winterhalter K. H., Zimmermann D. R. Brain derived versican V2 is a potent inhibitor of axonal growth. J Cell Sci. 2000 Mar;113(Pt 5):807–816. doi: 10.1242/jcs.113.5.807. [DOI] [PubMed] [Google Scholar]
  28. Schmalfeldt M., Dours-Zimmermann M. T., Winterhalter K. H., Zimmermann D. R. Versican V2 is a major extracellular matrix component of the mature bovine brain. J Biol Chem. 1998 Jun 19;273(25):15758–15764. doi: 10.1074/jbc.273.25.15758. [DOI] [PubMed] [Google Scholar]
  29. Westling Jennifer, Fosang Amanda J., Last Karena, Thompson Vivian P., Tomkinson Kathy N., Hebert Tracy, McDonagh Thomas, Collins-Racie Lisa A., LaVallie Edward R., Morris Elisabeth A. ADAMTS4 cleaves at the aggrecanase site (Glu373-Ala374) and secondarily at the matrix metalloproteinase site (Asn341-Phe342) in the aggrecan interglobular domain. J Biol Chem. 2002 Feb 19;277(18):16059–16066. doi: 10.1074/jbc.M108607200. [DOI] [PubMed] [Google Scholar]
  30. Yamada H., Watanabe K., Shimonaka M., Yamaguchi Y. Molecular cloning of brevican, a novel brain proteoglycan of the aggrecan/versican family. J Biol Chem. 1994 Apr 1;269(13):10119–10126. [PubMed] [Google Scholar]
  31. Yamaguchi Y. Lecticans: organizers of the brain extracellular matrix. Cell Mol Life Sci. 2000 Feb;57(2):276–289. doi: 10.1007/PL00000690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yuan W., Matthews R. T., Sandy J. D., Gottschall P. E. Association between protease-specific proteolytic cleavage of brevican and synaptic loss in the dentate gyrus of kainate-treated rats. Neuroscience. 2002;114(4):1091–1101. doi: 10.1016/s0306-4522(02)00347-0. [DOI] [PubMed] [Google Scholar]
  33. Zimmermann D. R., Dours-Zimmermann M. T., Schubert M., Bruckner-Tuderman L. Versican is expressed in the proliferating zone in the epidermis and in association with the elastic network of the dermis. J Cell Biol. 1994 Mar;124(5):817–825. doi: 10.1083/jcb.124.5.817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zimmermann D. R., Ruoslahti E. Multiple domains of the large fibroblast proteoglycan, versican. EMBO J. 1989 Oct;8(10):2975–2981. doi: 10.1002/j.1460-2075.1989.tb08447.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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