Kallunki P, Sainio K, Eddy R, Byers M, Kallunki T, Sariola H, Beck K, Hirvonen H, Shows T B, Tryggvason K
Department of Biochemistry, University of Oulu, Finland.
J Cell Biol. 1992 Nov;119(3):679-93. doi: 10.1083/jcb.119.3.679.
We describe the identification of a novel laminin chain. Overlapping clones were isolated from a human fibrosarcoma HT1080 cell cDNA library spanning a total of 5,200 bp. A second set of clones contained an alternative 3' end sequence giving a total of 4,316 bp. The longer sequence contained an open reading frame for a 1,193-residue-long polypeptide. The alternative sequence was shortened at the carboxyl-terminal end coding for a 1,111-residue-long polypeptide. The amino acid sequence contained 21 amino acids of a putative signal peptide and 1,172 residues or alternatively 1,090 residues of a sequence with five distinct domains homologous to domains I-V in laminin chains. Comparison of the amino acid sequences showed that the novel laminin chain is homologous to the laminin B2 chain. However, the structure of the novel laminin chain isolated here differs significantly from that of the B2 chain in that it has no domain VI and domains V, IV, and III are shorter, resulting in a truncated laminin chain. The alternative sequence had a shortened domain I/II. In accordance with the current nomenclature, the chain characterized here is termed B2t. Calculation of possible chain interactions of laminin chains with the B2t chain domain I/II indicated that the B2t chain can replace the B2 chain in some laminin molecules. The gene for the laminin B2t chain (LAMB2T) was localized to chromosome 1q25-q31 in close proximity to the laminin B2 chain gene. Northern analysis showed that the B2t chain is expressed in several human fetal tissues but differently from the laminin B1 and B2 chains. By in situ hybridization expression of the B2t chain was localized to specific epithelial cells in skin, lung, and kidney as opposed to a general epithelial and endothelial cell expression of the laminin B2 chain in the same tissues.
我们描述了一种新型层粘连蛋白链的鉴定过程。从人纤维肉瘤HT1080细胞cDNA文库中分离出重叠克隆,其总长为5200 bp。第二组克隆包含一个不同的3'端序列,总长为4316 bp。较长的序列含有一个编码1193个残基的多肽的开放阅读框。另一个序列在编码1111个残基的多肽的羧基末端被缩短。氨基酸序列包含21个氨基酸的假定信号肽以及1172个残基,或者是具有与层粘连蛋白链中I-V结构域同源的五个不同结构域的序列中的1090个残基。氨基酸序列比较表明,这种新型层粘连蛋白链与层粘连蛋白B2链同源。然而,这里分离出的新型层粘连蛋白链的结构与B2链有显著差异,因为它没有结构域VI,并且结构域V、IV和III较短,导致层粘连蛋白链被截断。另一个序列的结构域I/II被缩短。根据目前的命名法,这里鉴定的链被称为B2t。层粘连蛋白链与B2t链结构域I/II可能的链间相互作用计算表明,B2t链可以在一些层粘连蛋白分子中取代B2链。层粘连蛋白B2t链(LAMB2T)的基因定位于染色体1q25-q31,与层粘连蛋白B2链基因紧密相邻。Northern分析表明,B2t链在几种人类胎儿组织中表达,但与层粘连蛋白B1和B2链的表达不同。通过原位杂交,B2t链的表达定位于皮肤、肺和肾脏中的特定上皮细胞,而层粘连蛋白B2链在相同组织中是在上皮细胞和内皮细胞中普遍表达。