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首页> 外文期刊>Molecular and Cellular Biology >Characterization and cloning of a receptor for BMP-2 and BMP-4 from NIH 3T3 cells.
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Characterization and cloning of a receptor for BMP-2 and BMP-4 from NIH 3T3 cells.

机译：NIH 3T3细胞中BMP-2和BMP-4受体的表征和克隆。

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The bone morphogenetic proteins (BMPs) are a group of transforming growth factor beta (TGF-beta)-related factors whose only receptor identified to date is the product of the daf-4 gene from Caenorhabditis elegans. Mouse embryonic NIH 3T3 fibroblasts display high-affinity 125I-BMP-4 binding sites. Binding assays are not possible with the isoform 125I-BMP-2 unless the positively charged N-terminal sequence is removed to create a modified BMP-2, 125I-DR-BMP-2. Cross-competition experiments reveal that BMP-2 and BMP-4 interact with the same binding sites. Affinity cross-linking assays show that both BMPs interact with cell surface proteins corresponding in size to the type I (57- to 62-kDa) and type II (75- to 82-kDa) receptor components for TGF-beta and activin. Using a PCR approach, we have cloned a cDNA from NIH 3T3 cells which encodes a novel member of the transmembrane serine/threonine kinase family most closely resembling the cloned type I receptors for TGF-beta and activin. Transient expression of this receptor in COS-7 cells leads to an increase in specific 125I-BMP-4 binding and the appearance of a major affinity-labeled product of approximately 64 kDa that can be labeled by either tracer. This receptor has been named BRK-1 in recognition of its ability to bind BMP-2 and BMP-4 and its receptor kinase structure. Although BRK-1 does not require cotransfection of a type II receptor in order to bind ligand in COS cells, complex formation between BRK-1 and the BMP type II receptor DAF-4 can be demonstrated when the two receptors are coexpressed, affinity labeled, and immunoprecipitated with antibodies to either receptor subunit. We conclude that BRK-1 is a putative BMP type I receptor capable of interacting with a known type II receptor for BMPs.
机译：骨形态发生蛋白（BMP）是一组与转化生长因子β（TGF-β）相关的因子，迄今为止，其唯一鉴定出的受体是秀丽隐杆线虫的daf-4基因的产物。小鼠胚胎NIH 3T3成纤维细胞显示出高亲和力的125I-BMP-4结合位点。除非去除带正电的N端序列以生成修饰的BMP-2、125I-DR-BMP-2，否则同工型125I-BMP-2不可能进行结合测定。交叉竞争实验表明BMP-2和BMP-4与相同的结合位点相互作用。亲和力交联测定表明，两种BMP都与细胞表面蛋白相互作用，其大小对应于TGF-β和激活素的I型（57-62kDa）和II型（75-82kDa）受体组分。使用PCR方法，我们从NIH 3T3细胞克隆了cDNA，该cDNA编码跨膜丝氨酸/苏氨酸激酶家族的一个新成员，与TGF-β和激活素的克隆I型受体最相似。该受体在COS-7细胞中的瞬时表达导致特异性125I-BMP-4结合的增加以及大约64 kDa的主要亲和标记产物的出现，该产物可以被任一示踪剂标记。该受体因结合BMP-2和BMP-4的能力及其受体激酶结构而被命名为BRK-1。尽管BRK-1不需要共转染II型受体才能结合COS细胞中的配体，但是当两种受体共表达，亲和标记，BRK-1和BMP II型受体DAF-4之间可以形成复合物时，并用针对任一受体亚基的抗体进行免疫沉淀。我们得出结论，BRK-1是一种推定的BMP I型受体，能够与BMPs的已知II型受体相互作用。

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