Pilar Fernandez M, Selmin O, Martin G R, Yamada Y, Pfäffle M, Deutzmann R, Mollenhauer J, von der Mark K
Laboratory of Developmental Biology and Anomalies, National Institute of Dental Research, Bethesda, Maryland 20892.
J Biol Chem. 1988 Apr 25;263(12):5921-5.
cDNA clones for anchorin CII (Mr = 34,000), a collagen-binding protein, were isolated from a lambda gt 11 cDNA library prepared from chick cartilage mRNA. Several overlapping clones were characterized which gave rise to an open reading frame coding for 329 residues and a 3'-untranslated segment of 500 base pairs. The clones were identified as coding for anchorin by hybrid select translation analysis and by comparing the deduced amino acid sequence with the sequence of 10 tryptic peptides of the protein. A hydrophobic domain of 25 residues interrupted with 3 polar residues was identified with the carboxyl-terminal portion. There was no evidence for an aminoterminal signal peptide. Northern analysis revealed that the 5' probe hybridizes to a single 1.7-kilobases (kb) mRNA species, whereas the 3' probe hybridizes to two mRNA species of 1.7 kb and 5 kb, which are present in many cells including chondrocytes, crop cells, and fibroblasts. The level of anchorin mRNA in chick embryo fibroblasts was increased by infection with Rous sarcoma virus.
从用鸡软骨mRNA构建的λgt 11 cDNA文库中分离出锚定蛋白CII（分子量为34,000）的cDNA克隆，该蛋白是一种胶原结合蛋白。对几个重叠克隆进行了表征，它们产生了一个编码329个残基的开放阅读框和一个500个碱基对的3'非翻译区。通过杂交选择翻译分析以及将推导的氨基酸序列与该蛋白的10个胰蛋白酶肽段序列进行比较，确定这些克隆编码锚定蛋白。在羧基末端部分鉴定出一个由25个残基组成的疏水结构域，其中间有3个极性残基。没有证据表明存在氨基末端信号肽。Northern分析显示，5'探针与一种单一的1.7千碱基（kb）mRNA杂交，而3'探针与1.7 kb和5 kb的两种mRNA杂交，这两种mRNA存在于包括软骨细胞、嗉囊细胞和成纤维细胞在内的许多细胞中。用劳氏肉瘤病毒感染鸡胚成纤维细胞后，锚定蛋白mRNA的水平升高。
