Erle D J, Sheppard D, Breuss J, Rüegg C, Pytela R
Lung Biology Center, University of California, San Francisco 94143.
Am J Respir Cell Mol Biol. 1991 Aug;5(2):170-7. doi: 10.1165/ajrcmb/5.2.170.
The integrins are a large group of cell surface glycoproteins that mediate cell-matrix and cell-cell adhesive interactions. Integrins play a role in normal lung development, in host defense against pulmonary infection, and in the pathogenesis of the adult respiratory distress syndrome. Integrins are heterodimers consisting of one alpha subunit and one beta subunit. We identified consensus sequences within integrin subunits and used oligonucleotide primers based on these sequences to amplify cDNA by the polymerase chain reaction (PCR). We previously reported the use of this homology PCR technique for the identification of one novel integrin beta subunit, beta 6, from guinea pig airway epithelial cells. Here we demonstrate that primers based on alpha subunit consensus sequences can also be used for homology PCR. We have used the alpha and beta subunit primers to amplify and clone a large variety of integrin partial cDNAs from several cell types and species. Comparison of the deduced amino acid sequences reveals a high degree of cross-species conservation (86 to 98% identity). One alpha subunit (identified in guinea pig airway epithelial cells) and one beta subunit (identified in rabbit leukocytes obtained by bronchoalveolar lavage and in human and mouse leukocyte cell lines) have novel sequences that are related to but clearly distinct from all previously reported integrin sequences (24 to 61% identity). These novel cDNAs are very likely to encode previously unsequenced integrin subunit proteins that are expressed in the lung. Homology PCR is a powerful technique for the identification of known and novel integrin alpha and beta subunit cDNAs in cells from the lung and other organs.
整合素是一大类细胞表面糖蛋白,介导细胞与基质以及细胞与细胞之间的黏附相互作用。整合素在正常肺发育、宿主抵御肺部感染以及成人呼吸窘迫综合征的发病机制中发挥作用。整合素是由一个α亚基和一个β亚基组成的异二聚体。我们在整合素亚基中鉴定出共有序列,并基于这些序列使用寡核苷酸引物通过聚合酶链反应(PCR)扩增cDNA。我们之前报道过利用这种同源PCR技术从豚鼠气道上皮细胞中鉴定出一种新的整合素β亚基β6。在此我们证明基于α亚基共有序列的引物也可用于同源PCR。我们已使用α亚基和β亚基引物从多种细胞类型和物种中扩增并克隆了多种整合素部分cDNA。推导的氨基酸序列比较显示出高度的跨物种保守性(同一性为86%至98%)。一个α亚基(在豚鼠气道上皮细胞中鉴定出)和一个β亚基(在通过支气管肺泡灌洗获得的兔白细胞以及人和小鼠白细胞细胞系中鉴定出)具有与所有先前报道的整合素序列相关但明显不同的新序列(同一性为24%至61%)。这些新的cDNA很可能编码在肺中表达的先前未测序的整合素亚基蛋白。同源PCR是一种用于鉴定肺和其他器官细胞中已知和新的整合素α和β亚基cDNA的强大技术。