Polydispersity and heterogeneity of squid cranial cartilage proteoglycans as assessed by immunochemical methods and electron microscopy.
The three populations of squid cranial cartilage proteoglycans, D1D1A, D1D1B and D1D2 appeared to have a high degree of polydispersity. Gel electrophoresis and immunoblotting analysis showed that polydispersity was mainly due to the variable size of chondroitin sulphate E chains. This was further ascertained after rotary shadowing electron microscopy of proteoglycan core proteins and glycosaminoglycan side chains and statistical analysis of the sizes measured for both components. Enzymic treatment of the proteoglycan core proteins produced different peptides from each population, suggesting that the observed heterogeneity of the proteoglycans is due to their core proteins. Antibodies were raised in rabbits against all proteoglycans and enzyme-linked immunosorbent analysis of proteoglycan core proteins revealed that the proteoglycans, even heterogeneous, shared many common epitopes. Part of the common proteoglycan epitopes were found to be located in chondroitin sulphate E chains. Heterogeneity of squid proteoglycans was also investigated by studying their interactions with collagen and it was found that only the two populations of high molecular mass, D1D1A and D1D2, were able to interact with only collagen type I, the latter stronger than the former.
Vynios
D H
DH
Laboratory of Biochemistry, Department of Chemistry, University of Patras, 261 10, Patras, Greece. vynios@upatras.gr
Mörgelin
M
M
Papageorgakopoulou
N
N
Tsilemou
A
A
Spyracopoulou
G
G
Zafira
M
M
Tsiganos
C P
CP
D000818:Animals; D015153:Blotting, Western; D002356:Cartilage / Q000737:chemistry*; D019765:Chondroitin ABC Lyase; D049832:Decapodiformes; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000737:chemistry* / Q000648:ultrastructure*
D011509:Proteoglycans; D019765:Chondroitin ABC Lyase