G3 domains of aggrecan and PG-M/versican form intermolecular disulfide bonds that stabilize cell-matrix interaction.
The extracellular matrix plays a critical role in maintaining tissue integrity. Among the matrix molecules, the large aggregating chondroitin sulfate proteoglycans are the major structural molecules and are the primary contributors to the stability for some tissues such as cartilage. The notable exceptions are nanomelic cartilage and arthritic cartilage: the former contains a point mutation leading to a stop codon before translating to the C-terminal G3 domain; the latter contains a large proportion of aggrecan from which the G3 domain has been cleaved. These phenomena suggest that the G3 domain may be important in cartilage stability. Here, we demonstrated for the first time that the G3 domains of aggrecan and another proteoglycan, PG-M/versican, formed intermolecular disulfide bonds, and all subdomains were involved. Further studies indicated that each of the 10 cysteine residues of the aggrecan G3 domain could potentially form intermolecular disulfide bonds in vitro. The disulfide bonds were disrupted in the presence of reducing reagent beta-mercaptoethanol and dithiothreitol. As a result, normal chondrocyte-matrix interaction was disrupted, and the structure of the extracellular matrix was altered. Furthermore, disruption of disulfide bonds also reduced the role of PG-M/versican G3 domain in mediating cell adhesion. Our study provides strong evidence of the importance of proteoglycan interactions through intermolecular disulfide bonds in cartilage firmness and cell-matrix stability.
Chen
Liwen
L
Sunnybrook and Women's College Health Science, 2075 Bayview Avenue, Toronto M4N 3M5, Canada.
Yang
Bing L
BL
Wu
Yaojiong
Y
Yee
Albert
A
Yang
Burton B
BB
D053674:Aggrecans; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D003470:Culture Media; D017931:DNA Primers; D004220:Disulfides / Q000737:chemistry*; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011509:Proteoglycans / Q000737:chemistry*
D053674:Aggrecans; D003470:Culture Media; D017931:DNA Primers; D004220:Disulfides; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans