Processing of type II procollagen amino propeptide by matrix metalloproteinases
Processing of type II procollagen amino propeptide by matrix metalloproteinases
复制标题
DOI：
10.1074/jbc.m105485200
复制
发表时间：
2002-01-18
影响因子：
4.8
通讯作者：
Sandell, LJ
中科院分区：
文献类型：
--
作者：
Fukui, N;McAlinden, A;Sandell, LJ
在许多胚胎组织中，IIA型前胶原被合成并沉积到含有NH 2-前肽的细胞外基质中，NH 2-前肽的富含半胱氨酸的结构域与骨形态发生蛋白结合。为了研究在发育和疾病过程中合成的基质金属蛋白酶（MMPs）是否可以切割11型前胶原的NH 2末端，我们测试了8种酶。使用与大鼠表面活性蛋白D的凝集素结构域融合的外显子1-8编码的重组三聚体IIA型胶原NH 2前肽作为底物。后者允许前肽结构域的三聚化，并允许通过糖亲和色谱分离。虽然NEAP 1，2和8没有显示切割，MMPs 3，7，9，13和14切割重组蛋白在端肽区和前胶原N-蛋白酶切割位点。MMP 7和13在N-前肽的11型胶原特异性区域中表现出其他切割位点; MMP-7在靠近富含半胱氨酸的结构域的COOH末端处具有另一个切割位点。为了证明MMP可以原位切割天然IIA型前胶原，我们证明了MMP-7从胎儿软骨的细胞外基质中的胶原纤维中去除NH 2-前肽，并鉴定了切割产物。因为N-蛋白酶和端肽酶切割位点存在于IIA型和IIB型前胶原中，并且端肽切割位点保留在成熟胶原原纤维中，所以这种加工对IIB型前胶原和成熟胶原原纤维也是重要的。
in many embryonic tissues, type IIA procollagen is synthesized and deposited into the extracellular matrix containing the NH2-propeptide, the cysteine-rich domain of which binds to bone morphogenic proteins. To investigate whether matrix metalloproteinases (MMPs) synthesized during development and disease can cleave the NH2 terminus of type 11 procollagens, we tested eight types of enzymes. Recombinant trimeric type IIA collagen NH2-propeptide encoded by exons 1-8 fused to the lectin domain of rat surfactant protein D was used as a substrate. The latter allowed trimerization of the propeptide domain and permitted isolation by saccharide affinity chromatography. Although NEAPs 1, 2, and 8 did not show cleavage, MMPs 3, 7, 9, 13, and 14 cleaved the recombinant protein both at the telopeptide region and at the procollagen N-proteinase cleavage site. MMPs 7 and 13 demonstrated other cleavage sites in the type 11 collagen-specific region of the N-propeptide; MMP-7 had another cleavage site close to the COOH terminus of the cysteine-rich domain. To prove that an I MP can cleave the native type IIA procollagen in situ, we demonstrated that MMP-7 removes the NH2-propeptide from collagen fibrils in the extracellular matrix of fetal cartilage and identified the cleavage products. Because the N-proteinase and telopeptidase cleavage sites are present in both type IIA and type IIB procollagens and the telopeptide cleavage site is retained in the mature collagen fibril, this processing could be important to type IIB procollagen and to mature collagen fibrils as well.