Proteoglycans from the swarm rat chondrosarcoma. Structure of the aggregates extracted with associative and dissociative solvents as revealed by electron microscopy.
Proteoglycan aggregates were extracted from Swarm rat chondrosarcoma tissue in the native state and compared with proteoglycan aggregates isolated dissociatively with 4 M guanidine HCl. Purified aggregates were examined with a variety of electron microscopic techniques. In some cases they showed a structure of the central filament identical to that of the link-stabilized central filament observed in earlier experiments where the separated constituents were allowed to reconstitute (Mörgelin, M., Paulsson, M., Hardingham, T. E., Heinegård, D., and Engel, J. (1988) Biochem. J. 253, 175-185). The tight packing of proteoglycan monomers along the hyaluronate with a minimum distance of 12 nm between adjacent E1 strands also could thus be confirmed for never dissociated aggregates. The results therefore show that the organization of proteoglycan aggregates assembled in vitro from the participating molecules is representative for conditions in situ. An additional structural type of central filament was observed in the preparations. This contained long stretches of free hyaluronate interspaced by short stretches of central filament with condensed arrays of link protein-proteoglycan. Chemical cross-linking in combination with low shear electron microscopical techniques showed that this discontinuous central filament structure is not an artifact of specimen preparation. The addition of suprastoichiometric amounts of exogenous link protein did not affect the central filament structure with the low packing density. Densely and loosely packed types of central filament were isolated in varying relative amounts with different associative and dissociative solvents.
Mörgelin
M
M
M. E. Müller Institute for High Resolution Electron Microscopy, University of Basel, Switzerland.
Engel
J
J
Heinegård
D
D
Paulsson
M
M
D000818:Animals; D002240:Carbohydrate Sequence; D002810:Chondroitinases and Chondroitin Lyases; D002813:Chondrosarcoma / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D011509:Proteoglycans / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D051381:Rats; D011919:Rats, Inbred Strains; D012513:Sarcoma, Experimental / Q000737:chemistry*; D012997:Solvents
D003432:Cross-Linking Reagents; D009844:Oligosaccharides; D011509:Proteoglycans; D012997:Solvents; D002810:Chondroitinases and Chondroitin Lyases