Interaction between collagens and glycosaminoglycans investigated using a surface plasmon resonance biosensor

Abstract

The interactions of glycosaminoglycans with collagens and other glycoproteins in extracellular matrix play important roles in cell adhesion and extracellular matrix assembly. In order to clarify the chemical bases for these interactions, glycosaminoglycan solutions were injected onto sensor surfaces on which collagens, fibronectin, laminin, and vitronectin were immobilized. Heparin bound to type V collagen, type IX collagen, fibronectin, laminin, and vitronectin; and chondroitin sulfate E bound to type II, type V, and type VII collagen. Heparin showed a higher affinity for type IX collagen than for type V collagen. On the other hand, chondroitin sulfate E showed the highest affinity for type V collagen. The binding of chondroitin sulfate E to type V collagen showed higher affinity than that of heparin to type V collagen. These data suggest that a novel characteristic sequence included in chondroitin sulfate E is involved in binding to type V collagen.

Domain Structure of Chondroitin Sulfate E Octasaccharides Binding to Type V Collagen

“…The transglycosylation mechanism of testicular hyaluronidase, which is an endo-␤-N-acetylhexosaminidase, has been investigated with the aim of performing enzymatic synthesis of GAGs (9,10). It was possible to custom synthesize GAG oligosaccharides having various arrangements of nonsulfated disaccharide units (GlcA ␤1-3GalNAc, GlcA␤1-3GlcNAc, and IdoA␣1-3GalNAc) and monosulfated disaccharide units (GlcA␤1-3GalNAc(4S) and GlcA␤1-3GalNA(6S)), by repeating the transglycosylation using suitable combinations of HA, Ch, desulfated DS, Ch4S, and Ch6S as acceptors and donors.…”

“…Briefly, 50 g of one GAG (HA, Ch, Ch4S, Ch6S, or desulfated DS) as the donor, 20 nmol of a PA-oligosaccharide as the acceptor, and 5 units of testicular hyaluronidase dissolved in 50 g of 0.15 M Tris-HCl buffer, pH 7.0, were incubated at 37°C for 1 h. The reaction was terminated by immersion in a boiling water bath at 100°C for 3 min. The resulting product was purified by HPLC, and its structure was confirmed by a method described previously (9,10). Using systematic combinations of the donor and acceptor molecules, we prepared many chimeric type GAG oligosaccharides consisting of different combinations of disaccharide units (22,23).…”

“…Binding reactions caused a change in the surface plasmon resonance, which was detected optically and measured in resonance units. Immobilization of type V collagen on the sensor surface via primary amine groups was performed as described previously (9). The amounts of type V collagen immobilized onto the sensor surface were controlled at about 3000 resonance units by changing the injection time.…”

“…However, the relationship between the biological function and structure of ChS domains is not yet fully understood. Recently, we used a surface plasmon resonance (SPR) biosensor to investigate the interaction of glycosaminoglycans (GAGs) with collagens and glycoproteins from the extracellular matrix (9). It was found that chondroitin sulfate E (ChS-E) has specific affinity for type V collagen, and that a novel characteristic sequence included in ChS-E is probably involved in binding to type V collagen.…”

Structural Characterization of Heparan Sulfate and Chondroitin Sulfate of Syndecan-1 Purified from Normal Murine Mammary Gland Epithelial Cells

“…Similar regulatory mechanisms by the CS chains may function for other Hep-binding proteins, such as fibroblast growth factor-2 (67), morphogen-like Wingless and Hedgehog proteins (22), lipoprotein lipase (68), and various collagen types (69), which specifically bind to the HS chains of syndecan-1. Notably, CS-E has been shown to specifically bind to lipoprotein lipase (70) and multiple collagen types as well (71). DS chains that are structurally analogous to CS, released after injury, specifically bind to fibroblast growth factor-2 to promote its cell proliferation activity (72).…”