Molecular cloning of amphiglycan, a novel integral membrane heparan sulfate proteoglycan expressed by epithelial and fibroblastic cells.
Molecular cloning of amphiglycan, a novel integral membrane heparan sulfate proteoglycan expressed by epithelial and fibroblastic cells.
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DOI：
10.1083/jcb.118.4.961
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发表时间：
1992-08
期刊：
影响因子：
--
通讯作者：
van den Berghe H
中科院分区：
文献类型：
--
作者：
David G;van der Schueren B;Marynen P;Cassiman JJ;van den Berghe H
我们合成了一种反义寡核苷酸引物，该引物与具有跨膜方向的硫酸乙酰肝素蛋白聚糖的消息中的假定保守序列相匹配。借助该引物，我们扩增了来自人肺成纤维细胞的信息的部分和选定的全长拷贝，该信息编码新型整膜硫酸乙酰肝素蛋白多糖。编码的蛋白质长 198 个氨基酸，具有离散的细胞质、跨膜和氨基末端胞外结构域。除了代表假定的硫酸乙酰肝素链附着位点的序列外，该蛋白质的胞外结构域具有独特的结构。相反，跨膜和细胞质结构域与纤维聚糖和多聚糖的相应结构域高度相似，这两种细胞表面蛋白聚糖被视为反义引物设计的模型。这种相似性包括四个酪氨酸残基的保守性，一个紧邻终止转移序列前面，三个在细胞质片段中，以及最近端和最远端细胞质序列的保守性。该 cDNA 可检测培养的人肺成纤维细胞以及各种人上皮细胞和成纤维细胞系中的单个 2.6 kb 信息。在表达为 β-半乳糖苷酶融合蛋白后，针对所编码的肽产生的多克隆和单克隆抗体与人肺成纤维细胞的细胞表面硫酸乙酰肝素蛋白聚糖的 35-kD 核心蛋白反应，并装饰许多细胞类型的表面。我们建议将这种蛋白多糖命名为“amphiglycan”（来自希腊词amphi，“围绕，在两侧”和amphoo，“两者”），指的是其在质膜两侧延伸的结构域，以及其定位在上皮细胞和成纤维细胞来源的细胞周围。
We have synthesized an antisense oligonucleotide primer that matches a supposedly conserved sequence in messages for heparan sulfate proteoglycans with transmembrane orientations. With the aid of this primer we have amplified partial and selected full-length copies of a message from human lung fibroblasts that codes for a novel integral membrane heparan sulfate proteoglycan. The encoded protein is 198 amino- acids long, with discrete cytoplasmic, transmembrane, and amino- terminal extracellular domains. Except for the sequences that represent putative heparan sulfate chain attachment sites, the extracellular domain of this protein has a unique structure. The transmembrane and cytoplasmic domains, in contrast, are highly similar to the corresponding domains of fibroglycan and syndecan, the two cell surface proteoglycans that figured as models for the design of the antisense primer. This similarity includes the conservation of four tyrosine residues, one immediately in front of the stop transfer sequence and three in the cytoplasmic segment, and of the most proximal and most distal cytoplasmic sequences. The cDNA detects a single 2.6-kb message in cultured human lung fibroblasts and in a variety of human epithelial and fibroblastic cell lines. Polyclonal and monoclonal antibodies raised against the encoded peptide after expression as a beta- galactosidase fusion protein react with the 35-kD coreprotein of a cell surface heparan sulfate proteoglycan of human lung fibroblasts and decorate the surface of many cell types. We propose to name this proteoglycan "amphiglycan" (from the Greek words amphi, "around, on both sides of" and amphoo, "both") referring to its domain structure which extends on both sides of the plasmamembrane, and to its localization around cells of both epithelial and fibroblastic origin.