Myers J C, Kivirikko S, Gordon M K, Dion A S, Pihlajaniemi T
Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia 19104-6059.
Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10144-8. doi: 10.1073/pnas.89.21.10144.
A previously unknown collagen cDNA clone, PF19, was isolated from a human placenta library. The 2.1-kilobase insert has a complete open reading frame of 709 amino acids that includes 12 amino acids of the NH2-terminal domain, a principally collagenous region of 577 residues, and 120 residues of the noncollagenous COOH terminus. The collagenous part of the sequence encoded by PF19 is characterized by 13 interruptions ranging in size from 2 to 45 amino acids. Within four interruptions are consensus sequences for attachment of serine-linked glycosaminoglycans and asparagine-linked oligosaccharides suggesting that this collagen may be extensively glycosylated. A synthetic decapeptide representing a sequence at the beginning of the COOH-terminal noncollagenous domain was used to prepare an antibody in rabbits. This antiserum detected a 125-kDa bacterial collagenase-sensitive protein in Western blots of HeLa cell lysate. Consistent with the size of the collagen chain, Northern blot hybridization revealed a major transcript of 5.3 kilobases and two minor ones of 4.7 and 4.4 kilobases that are present in cultured human fibroblasts but absent from umbilical vein endothelial cells. We propose that the previously unidentified polypeptide described in this report be designated the alpha 1 chain of type XV collagen.
从人胎盘文库中分离出一个先前未知的胶原cDNA克隆PF19。2.1千碱基的插入片段有一个709个氨基酸的完整开放阅读框，其中包括12个氨基末端结构域的氨基酸、一个由577个残基组成的主要胶原区域以及120个非胶原羧基末端的残基。PF19编码序列的胶原部分的特征是有13个中断，大小从2到45个氨基酸不等。在四个中断处有丝氨酸连接的糖胺聚糖和天冬酰胺连接的寡糖附着的共有序列，这表明这种胶原可能被广泛糖基化。一个代表羧基末端非胶原结构域起始序列的合成十肽被用来在兔中制备抗体。该抗血清在HeLa细胞裂解物的蛋白质印迹中检测到一种125 kDa的对细菌胶原酶敏感的蛋白质。与胶原链的大小一致，Northern印迹杂交显示有一个5.3千碱基的主要转录本以及两个4.7和4.4千碱基的次要转录本存在于培养的人成纤维细胞中，而脐静脉内皮细胞中没有。我们建议将本报告中描述的先前未鉴定的多肽命名为XV型胶原的α1链。
