Piecha Dorothea, Wiberg Charlotte, Mörgelin Matthias, Reinhardt Dieter P, Deák Ferenc, Maurer Patrik, Paulsson Mats
Institute for Biochemistry, Medical Faculty, University of Cologne, Joseph-Stelzmann-Str. 52, D-50931 Cologne, Germany.
Biochem J. 2002 Nov 1;367(Pt 3):715-21. doi: 10.1042/BJ20021069.
Matrilin-2 is a component of extracellular filamentous networks. To study the interactions by which it can be integrated into such assemblies, full-length and truncated forms of matrilin-2 were recombinantly expressed in HEK-293 cells and purified from conditioned medium. The recombinant proteins, when used in interaction assays, showed affinity to matrilin-2 itself, but also to other collagenous and non-collagenous extracellular matrix proteins. The interaction between matrilin-2 and collagen I was studied in greater detail and could be shown to occur at distinct sites on the collagen I molecule and to have a K (D) of about 3 x 10(-8) M. Interactions with some non-collagenous protein ligands were even stronger, with matrilin-2 binding to fibrillin-2, fibronectin and laminin-1-nidogen-1 complexes, with K (D) values in the range of 10(-8)-10(-11) M. Co-localization of matrilin-2 with these ligands in the dermal-epidermal basement membrane, in the microfibrils extending from the basement membrane into the dermis, and in the dermal extracellular matrix, indicates a physiological relevance of the interactions in the assembly of supramolecular extracellular matrix structures.
基质金属蛋白酶-2是细胞外丝状网络的一个组成部分。为了研究其能够整合到此类组件中的相互作用，基质金属蛋白酶-2的全长和截短形式在人胚肾293细胞中进行重组表达，并从条件培养基中纯化。这些重组蛋白在相互作用分析中，不仅显示出对基质金属蛋白酶-2自身的亲和力，还显示出对其他胶原和非胶原细胞外基质蛋白的亲和力。对基质金属蛋白酶-2与I型胶原之间的相互作用进行了更详细的研究，结果表明这种相互作用发生在I型胶原分子的不同位点，解离常数（KD）约为3×10⁻⁸M。与一些非胶原蛋白配体的相互作用甚至更强，基质金属蛋白酶-2与原纤维蛋白-2、纤连蛋白以及层粘连蛋白-1-巢蛋白-1复合物结合，KD值在10⁻⁸ - 10⁻¹¹M范围内。基质金属蛋白酶-2与这些配体在真皮-表皮基底膜、从基底膜延伸至真皮的微纤维以及真皮细胞外基质中的共定位，表明这些相互作用在超分子细胞外基质结构组装中具有生理相关性。
