Calreticulin, PDI, Grp94 and BiP chaperone proteins are associated with retained COMP in pseudoachondroplasia chondrocytes

Matrix Biol. 2001 Jul;20(4):251-62. doi: 10.1016/s0945-053x(01)00136-6.

Abstract

Cartilage oligomeric matrix protein (COMP), a large pentameric glycoprotein and member of the thrombospondin (TSP) group of extracellular proteins, is found in the territorial matrix surrounding chondrocytes. More than 50 unique COMP mutations have been identified as causing two skeletal dysplasias: pseudoachondroplasia (PSACH); and multiple epiphyseal dysplasia (EDM1). Recent studies suggest that calcium-binding and calcium-induced protein folding differ between wild type and mutant proteins, and abnormal processing of the mutant COMP protein contributes to the characteristic enlarged lamellar appearing rER cisternae in PSACH and EDMI chondrocytes in vivo and in vitro. Towards the goal of delineating the pathogenesis of PSACH and EDM1, in-vivo PSACH growth plate and in-vitro PSACH chondrocytes cultured in alginate beads were examined to identify and localize the chaperone proteins participating in the processing of the retained extracellular matrix proteins in the PSACH rER. Aggrecan was localized to both the rER cisternae and matrix while COMP and type IX collagen were only found in the rER. Type II collagen was solely found in the ECM suggesting that it is processed and transported differently from other retained ECM proteins. Five chaperone proteins: BiP (Grp78); calreticulin (CRT); protein disulfide (PDI); ERp72; and Grp94, demonstrated immunoreactivity in the enlarged PSACH cisternae and the short rER channels of chondrocytes from both in-vivo and in-vitro samples. The chaperone proteins cluster around the electron dense material within the enlarged rER cisternae. CRT, PDI and GRP94 AB-gold particles appear to be closely associated with COMP. Immunoprecipitation and Western blot, and Fluorescence Resonance Energy Transfer (FRET) analyses indicate that CRT, PDI and GRP94 are in close proximity to normal and mutant COMP and BiP to mutant COMP. These results suggest that these proteins play a role in the processing and transport of wild type COMP in normal chondrocytes and in the retention of mutant COMP in PSACH chondrocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Achondroplasia / metabolism*
  • Achondroplasia / pathology
  • Calcium-Binding Proteins / metabolism*
  • Calreticulin
  • Carrier Proteins / metabolism*
  • Cartilage Oligomeric Matrix Protein
  • Chondrocytes / metabolism*
  • Collagen / metabolism
  • Endoplasmic Reticulum Chaperone BiP
  • Endoplasmic Reticulum, Rough / metabolism
  • Extracellular Matrix Proteins / metabolism*
  • Glycoproteins / metabolism*
  • HSP70 Heat-Shock Proteins / metabolism*
  • Heat-Shock Proteins*
  • Humans
  • Matrilin Proteins
  • Membrane Glycoproteins / metabolism
  • Membrane Proteins / metabolism*
  • Molecular Chaperones / metabolism*
  • Osteochondrodysplasias / metabolism*
  • Osteochondrodysplasias / pathology
  • Ribonucleoproteins / metabolism*

Substances

  • Calcium-Binding Proteins
  • Calreticulin
  • Carrier Proteins
  • Cartilage Oligomeric Matrix Protein
  • Endoplasmic Reticulum Chaperone BiP
  • Extracellular Matrix Proteins
  • Glycoproteins
  • HSP70 Heat-Shock Proteins
  • HSPA5 protein, human
  • Heat-Shock Proteins
  • Matrilin Proteins
  • Membrane Glycoproteins
  • Membrane Proteins
  • Molecular Chaperones
  • Ribonucleoproteins
  • TSP5 protein, human
  • endoplasmic reticulum glycoprotein p72
  • glucose-regulated proteins
  • Collagen