软骨胶原与肝素的相互作用

Gerald N. Smith Jr. , Kenneth D. Brandt
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引用次数: 20

摘要

软骨胶原在0.1 M NaCl/2 M尿素/0.01 M Tris-HC1溶液中与肝素-琼脂糖结合,可用线性梯度NaCl洗脱。洗脱峰出现在0.37 M NaCl处。变性的1 α和2α链结合强烈,这表明在这两条链上至少存在一个结合位点。软骨胶原除II型外,还包括几种次要的胶原类型(Burgeson和Hollister, 1979;Reese and Mayne, 1981)。这种结合在NaCl浓度增加时是稳定的,直到胶原原纤维开始溶解。肝素和硫酸葡聚糖是非常有效的相互作用抑制剂。如果是这样,这将提供一种方法来纯化软骨胶原,并评估多阴离子与软骨中各种胶原类型之间相互作用的相对强度。在本文中,我们展示了可溶性胶原与固定在Sepharose 4B上的肝素的结合,并检查了破坏这种结合所需的NaCl浓度。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Interaction of Cartilage Collagens with Heparin

Type XI collagen (1α2α3α) extracted from bovine articular cartilage by pepsin digestion binds strongly to heparin immobilized on agarose. The collagens from cartilage will bind to heparin-agarose in 0.1 M NaCl/2 M urea/0.01 M Tris-HC1 and can be eluted with a linear gradient of NaCl. Type XI begins to elute at NaCl concentrations higher than 0.28 M and is totally eluted at 0.40 M NaCl. The peak of elution occurs at 0.37 M NaCl. The other collagens of cartilage bind more weakly and are fully eluted when the NaCl concentration reaches 0.25 M. Collagen types I, III, and V are also bound to the column at low salt concentrations but are fully eluted before type XI begins to elute. All of the type XI preparation binds to heparin-agarose, suggesting that there is at least one binding site per molecule. Denatured 1 α and 2α chains bind strongly, suggesting that at least one binding site exists on both of these chains. These data confirm that the interaction between polyanions and type XI collagen is stronger than that with other known collagens and provide a method for purification of type XI without any type II contamination.

Cartilage collagens include several minor collagen types in addition to type II (Burgeson and Hollister, 1979; Reese and Mayne, 1981). Type XI, the collagen composed of the a chains designated lα, 2α, and 3α, binds to the high density proteoglycans of cartilage in vitro, apparently in a non-specific manner (Smith et al., 1985). This binding is stable in increasing concentrations of NaCl until the collagen fibrils begin to dissolve. Heparin and dextran sulfate are extremely potent inhibitors of the interaction.

The ability of heparin and dextran sulfate to inhibit the binding of fibrillar collagen to cartilage proteoglycans suggested that binding might also occur between type XI collagen and these highly sulfated polyanions. If so, this would provide a method to purify the cartilage collagens and to assess the relative strength of the interactions between the polyanion and the various collagen types in cartilage.

In this paper, we demonstrate the binding of soluble collagens to heparin immobilized on Sepharose 4B and examine the concentration of NaCl required to disrupt this binding.

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