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Determinants of versican-V1 proteoglycan processing by the metalloproteinase ADAMTS5.

  • Foulcer SJ From the Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
  • Nelson CM From the Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
  • Quintero MV the Departments of Medicinal Chemistry and Bioengineering, University of Utah Health Sciences Center, Salt Lake City, Utah 84112.
  • Kuberan B the Departments of Medicinal Chemistry and Bioengineering, University of Utah Health Sciences Center, Salt Lake City, Utah 84112.
  • Larkin J the Experimental Medicine Unit, GlaxoSmithKline, King of Prussia, Pennsylvania 19406, and.
  • Dours-Zimmermann MT the Institute of Surgical Pathology, University Hospital of Zurich, 8091 Zurich, Switzerland.
  • Zimmermann DR the Institute of Surgical Pathology, University Hospital of Zurich, 8091 Zurich, Switzerland.
  • Apte SS From the Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195, aptes@ccf.org.
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  • 2014-08-15
Published in:
  • The Journal of biological chemistry. - 2014
ADAMTS
Aggrecanase
Blocking Antibody
Chondroitin Sulfate
Extracellular Matrix
Glycosaminoglycan
Metalloprotease
Proteoglycan
Proteolytic Enzyme
Versican
ADAM Proteins
ADAMTS1 Protein
ADAMTS5 Protein
Amino Acid Motifs
Chondroitin Sulfates
Humans
Protein Binding
Protein Structure, Tertiary
Proteolysis
Versicans
English Proteolysis of the Glu(441)-Ala(442) bond in the glycosaminoglycan (GAG) β domain of the versican-V1 variant by a disintegrin-like and metalloproteinase domain with thrombospondin type 1 motif (ADAMTS) proteases is required for proper embryo morphogenesis. However, the processing mechanism and the possibility of additional ADAMTS-cleaved processing sites are unknown. We demonstrate here that if Glu(441) is mutated, ADAMTS5 cleaves inefficiently at a proximate upstream site but normally does not cleave elsewhere within the GAGβ domain. Chondroitin sulfate (CS) modification of versican is a prerequisite for cleavage at the Glu(441)-Ala(442) site, as demonstrated by reduced processing of CS-deficient or chondroitinase ABC-treated versican-V1. Site-directed mutagenesis identified the N-terminal CS attachment sites Ser(507) and Ser(525) as essential for processing of the Glu(441)-Ala(442) bond by ADAMTS5. A construct including only these two GAG chains, but not downstream GAG attachment sites, was cleaved efficiently. Therefore, CS chain attachment to Ser(507) and Ser(525) is necessary and sufficient for versican proteolysis by ADAMTS5. Mutagenesis of Glu(441) and an antibody to a peptide spanning Thr(432)-Gly(445) (i.e. containing the scissile bond) reduced versican-V1 processing. ADAMTS5 lacking the C-terminal ancillary domain did not cleave versican, and an ADAMTS5 ancillary domain construct bound versican-V1 via the CS chains. We conclude that docking of ADAMTS5 with two N-terminal GAG chains of versican-V1 via its ancillary domain is required for versican processing at Glu(441)-Ala(442). V1 proteolysis by ADAMTS1 demonstrated a similar requirement for the N-terminal GAG chains and Glu(441). Therefore, versican cleavage can be inhibited substantially by mutation of Glu(441), Ser(507), and Ser(525) or by an antibody to the region of the scissile bond.
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  • English
Open access status
bronze
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https://sonar.ch/global/documents/161193
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