PRIMARY STRUCTURE OF AN EXTRACELLULAR-MATRIX PROTEOGLYCAN CORE PROTEIN DEDUCED FROM CLONED CDNA
PRIMARY STRUCTURE OF AN EXTRACELLULAR-MATRIX PROTEOGLYCAN CORE PROTEIN DEDUCED FROM CLONED CDNA
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DOI：
10.1073/pnas.83.20.7683
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发表时间：
1986-10-01
影响因子：
11.1
通讯作者：
RUOSLAHTI, E
中科院分区：
文献类型：
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作者：
KRUSIUS, T;RUOSLAHTI, E
由人成纤维细胞表达并存在于细胞外基质中与胶原蛋白结合的小软骨素/硫酸皮肤素蛋白聚糖的核心蛋白已从A.gt11成纤维细胞cDNA文库中克隆。通过使用对完整蛋白聚糖特异的抗体和针对基于核心蛋白的氨基末端序列合成的肽的抗体分离cDNA克隆。发现1.8-腺苷酸酶cDNA编码由信号肽、前肽和329个氨基酸的成熟核心蛋白组成的前原核心蛋白。从cDNA序列推导的氨基末端氨基酸序列与先前通过蛋白质测序获得的氨基酸序列相同。核心蛋白含有三个Ser-Gly二肽序列，其中一个被糖胺聚糖取代。蛋白质数据库同源性搜索确定核心蛋白质序列是不同于已发表的氨基酸序列的独特序列。RNA印迹杂交，使用克隆的cDNA作为探针进行，揭示了两个相关的转录本1.6和1.9的RNA从人成纤维细胞和胎盘组织。基因组DNA限制性片段的杂交表明，有一个基因的核心蛋白，这种蛋白多糖和可能的其他密切相关的基因。蛋白聚糖的克隆cDNA的可用性现在使得有可能应用分子生物学方法来研究这种蛋白聚糖的胶原结合和细胞附着抑制特性。
The core protein of a small chondroitin/dermatan sulfate proteoglycan expressed by human fibroblasts and present in extracellular matrices in association with collagen has been cloned from a .lambda.gt11 fibroblast cDNA library. cDNA clones were isolated by use of antibodies specific for the intact proteoglycan and antibodies against a peptide synthesized on the basis of the amino-terminal sequence of the core protein. A 1.8-kilobase cDNA was found to code for a prepro core protein composed of a signal peptide, a propeptide, and a mature core protein of 329 amino acids. The amino-terminal amino acid sequence deduced from the cDNA sequence was identical to that previously obtained by protein sequencing. The core protein contains three Ser-Gly dipeptide sequences, of which one is substituted with glycosaminoglycan. A protein data base homology search established the core protein sequence is a unique sequence distinct from published amino acid sequences. RNA blot hybrdizations, performed using the cloned cDNA as a probe, revealed two related transcripts of 1.6 and 1.9 kilobases in RNA from both human fibroblast and placental tissue. Hybridization of genomic DNA restriction fragments suggested that there is one gene for the core protein of this proteoglycan and possibly one other closely related gene. Availability of the cloned cDNA for the proteoglycan now makes it possible to apply methods of molecular biology to study the collagen-binding and cell attachment-inhibiting properties of this proteoglycan.