title: Glycosaminoglycan polysulfate-induced stimulation of hyaluronic acid synthesis in rabbit knee synovial membrane: involvement of binding protein and calcium ion.
abstract: We have previously reported that naturally occurring sulfated glycosaminoglycans having a chondroitin-type structure and glycosaminoglycan polysulfate (GAGPS, a persulfated derivative of chondroitin sulfate) caused a specific stimulation of hyaluronic acid synthesis in rabbit knee synovial membranes [H. Nishikawa et al. (1985) Arch. Biochem. Biophys. 240, 146-153]. In the present study, the interaction of [3H]GAGPS and the surface of the rabbit knee synovial membranes and the relationship between this interaction and the stimulatory effect of GAGPS on the hyaluronic acid synthesis were examined in order to define the stimulatory mechanism of hyaluronic acid synthesis by GAGPS. A part of the [3H]GAGPS taken up by the synovial membranes was released from the membranes by trypsin treatment. The interaction of [3H]GAGPS and the surface of the isolated synovial membranes was diminished by pretreatment of the membranes with proteases or chelating reagents. Pretreatment of synovial membranes with trypsin or ethylene glycol bis(beta-aminoethyl ether) N,N'-tetraacetic acid had little effect on the basal hyaluronic acid synthesis but caused the loss of GAGPS-induced stimulation of hyaluronic acid synthesis accompanied by significant decrease (20% P less than 0.05-P less than 0.01) in the interaction between GAGPS and the surface of the synovial membranes. Dermatan sulfate having a chondroitin-type structure also stimulated hyaluronic acid synthesis but this effect was not additive to the stimulation of hyaluronic acid synthesis by GAGPS. Heparin had no effect on either the basal hyaluronic acid synthesis or the GAGPS-induced stimulation of hyaluronic acid synthesis. These results indicate that binding of GAGPS to certain distinct protein components on the surface of synovial membranes is involved in the stimulatory mechanism of hyaluronic acid synthesis by GAGPS, and that the binding may be mediated by Ca2+ ion. The binding was also found to be specific for sulfated glycosaminoglycans having a chondroitin-type structure.
authors.affiliation: Department of Pharmaceutical Radio Chemistry, Kobe Women's College of Pharmacy, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Nishikawa
authors.forename: I
authors.initials: I
authors.lastname: Mori
authors.forename: J
authors.initials: J
authors.lastname: Umemoto
mesh_terms: D000818:Animals; D002118:Calcium / Q000502:physiology*; D006025:Glycosaminoglycans / Q000378:metabolism / Q000494:pharmacology*; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D066298:In Vitro Techniques; D011485:Protein Binding; D011817:Rabbits; D013499:Surface Properties; D013583:Synovial Membrane / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
chemical_list: D006025:Glycosaminoglycans; C013786:A73025; D006820:Hyaluronic Acid; D014357:Trypsin; D002118:Calcium