Watanabe K, Yamada H, Yamaguchi Y
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037, USA.
J Cell Biol. 1995 Sep;130(5):1207-18. doi: 10.1083/jcb.130.5.1207.
Glypicans are a family of glycosylphosphatidylinositol (GPI)-anchored cell surface heparan sulfate proteoglycans (HSPGs). The glypican family, which currently includes glypican, the developmentally regulated rat intestinal transcript OCI-5, and cerebroglycan, is characterized by a similar core protein size and almost complete conservation of cysteine residues. By RT-PCR using degenerate oligonucleotide primers based on the sequence homologies, we isolated mouse cDNA encoding a novel member of the glypican family as well as mouse homologues of glypican and OCI-5. The novel molecule, named K-glypican, has a predicted molecular mass of 57.5 kD and potential attachment sites for heparan sulfate chains and a GPI anchor in its COOH-terminal region, like other members of the glypican family. Transfection of an epitope-tagged full-length K-glypican cDNA into MDCK cells demonstrated that K-glypican is indeed expressed as a GPI-anchored HSPG. Northern blot analyses with K-glypican, glypican, and OCI-5 probes demonstrated that kidney and developing brain, and that these three molecules show remarkable patterns of cell type- and developmental stage-specific expression. In situ hybridization revealed that the major sites of K-glypican expression in developing embryo are tubular epithelial cells in the kidney and proliferating neuroepithelial cells in the brain. These results indicate that K-glypican is a novel GPI-anchored HSPG involved in embryonic development.
磷脂酰肌醇蛋白聚糖是一类糖基磷脂酰肌醇(GPI)锚定的细胞表面硫酸乙酰肝素蛋白聚糖(HSPG)。磷脂酰肌醇蛋白聚糖家族目前包括磷脂酰肌醇蛋白聚糖、发育调控的大鼠肠道转录本OCI-5和脑硫酸乙酰肝素蛋白聚糖,其特征是核心蛋白大小相似且半胱氨酸残基几乎完全保守。通过使用基于序列同源性的简并寡核苷酸引物进行逆转录聚合酶链反应(RT-PCR),我们分离出了编码磷脂酰肌醇蛋白聚糖家族一个新成员以及磷脂酰肌醇蛋白聚糖和OCI-5的小鼠同源物的小鼠互补DNA(cDNA)。这个新分子名为K-磷脂酰肌醇蛋白聚糖,预测分子量为57.5kD,在其COOH末端区域有硫酸乙酰肝素链和GPI锚的潜在附着位点,与磷脂酰肌醇蛋白聚糖家族的其他成员一样。将带有表位标签的全长K-磷脂酰肌醇蛋白聚糖cDNA转染到MDCK细胞中表明,K-磷脂酰肌醇蛋白聚糖确实作为一种GPI锚定的HSPG表达。用K-磷脂酰肌醇蛋白聚糖、磷脂酰肌醇蛋白聚糖和OCI-5探针进行的Northern印迹分析表明,肾脏和发育中的脑表达这些分子,并且这三种分子显示出细胞类型和发育阶段特异性表达的显著模式。原位杂交显示,发育胚胎中K-磷脂酰肌醇蛋白聚糖表达的主要部位是肾脏中的肾小管上皮细胞和脑中增殖的神经上皮细胞。这些结果表明,K-磷脂酰肌醇蛋白聚糖是一种参与胚胎发育的新型GPI锚定HSPG。