Identification of cDNA clones encoding different domains of the basement membrane heparan sulfate proteoglycan.
Identification of cDNA clones encoding different domains of the basement membrane heparan sulfate proteoglycan.
复制标题
编码基底膜硫酸乙酰肝素蛋白多糖不同结构域的 cDNA 克隆的鉴定。
DOI：
10.1016/s0021-9258(18)37604-x
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
John R. Hassell
中科院分区：
文献类型：
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作者：
Douglas M. Noonan;E. Horigan;Steven R. Ledbetter;G. Vogeli;Makoto Sasaki;Y. Yamada;John R. Hassell
我们使用基底膜蛋白多糖抗体来筛选 lambda gt11 表达载体文库，并分离出两个 cDNA 克隆，称为 BPG 5 和 BPG 7，它们编码硫酸乙酰肝素基底膜蛋白多糖核心蛋白的不同部分。这些克隆与大约 12 kbps 的单一 mRNA 物种杂交。在推导的序列中发现了从核心蛋白的蛋白酶消化衍生的肽上获得的氨基酸序列，证实了这些克隆的身份。 BPG 5 跨越 1986 个碱基对，具有 662 个氨基酸的开放阅读框。从BPG 5推导的氨基酸序列包含两个富含半胱氨酸的结构域和两个缺乏半胱氨酸的内部同源结构域。富含半胱氨酸的结构域与层粘连蛋白链的富含半胱氨酸的结构域表现出同源性。蛋白多糖的该区域被提议采用与层粘连蛋白链的短臂相似的球杆结构。另一个克隆 BPG 7 长 2193 个碱基对，具有 731 个氨基酸的开放阅读框。推导的序列包含八个内部重复序列，每个重复序列中有 2 个半胱氨酸残基。这些重复序列与神经细胞粘附分子 N-CAM 和血浆 α1B-糖蛋白具有同源性。提出了与这些蛋白和免疫球蛋白基因超家族的其他蛋白类似的环结构用于蛋白多糖的这个区域。序列 DSGEY 在此结构域中被发现四次，可能是硫酸乙酰肝素附着位点。
We have used antibodies to the basement membrane proteoglycan to screen lambda gt11 expression vector libraries and have isolated two cDNA clones, termed BPG 5 and BPG 7, which encode different portions of the core protein of the heparan sulfate basement membrane proteoglycan. These clones hybridize to a single mRNA species of approximately 12 kilobases. Amino acid sequences obtained on peptides derived from protease digests of the core protein were found in the deduced sequence, confirming the identity of these clones. BPG 5 spanned 1986 base pairs and has an open reading frame of 662 amino acids. The amino acid sequence deduced from BPG 5 contains two cysteine-rich domains and two internally homologous domains lacking cysteine. The cysteine-rich domains show homology to the cysteine-rich domains of the laminin chains. A globule-rod structure, similar to that of the short arms of the laminin chains, is proposed for this region of the proteoglycan. The other clone, BPG 7, is 2193 base pairs long and has an open reading frame of 731 amino acids. The deduced sequence contains eight internal repeats with 2 cysteine residues in each repeat. These repeats show homology to the neural-cell adhesion molecule N-CAM and the plasma alpha 1B-glycoprotein. Looping structures similar to these proteins and to other proteins of the immunoglobulin gene superfamily are proposed for this region of the proteoglycan. The sequence DSGEY was found four times in this domain and could be heparan sulfate attachment sites.