Kayyem J F, Roman J M, de la Rosa E J, Schwarz U, Dreyer W J
California Institute of Technology, Pasadena 91125.
J Cell Biol. 1992 Sep;118(5):1259-70. doi: 10.1083/jcb.118.5.1259.
Diverse cell-surface molecules of the nervous system play an important role in specifying cell interactions during development. Using a method designed to generate mAbs against neural surface molecules of defined molecular weight, we have previously reported on the surface protein, Bravo, found in the developing avian retinotectal system. Bravo is immunologically detected on developing optic fibers in the retina, but absent from distal regions of the same fibers in the tectum. We have isolated cDNA clones encompassing the entire coding region of Bravo, including clones containing five alternative sequences of cDNA. These putative alternatively spliced sequences encode stretches of polypeptide ranging in length from 10-93 amino acids and are predicted to be both extra- and intracellular. The deduced primary structure of Bravo reveals that, like the cell adhesion molecules (CAMs) chicken Ng-CAM and mouse L1, Bravo is composed of six Ig-like domains, five fibronectin type III repeats, a transmembrane domain, and a short cytoplasmic region. Recently, the cDNA sequence of a related molecule, Nr-CAM, was reported and its possible identity with Bravo discussed (Grumet, M., V. Mauro, M. P. Burgoon, G. E. Edelman, and B. A. Cunningham. 1991. J. Cell Biol. 113:1399-1412). Here we confirm this identity and moreover show that Bravo is found on Müller glial processes and end-feet in the developing retina. In contrast to the single polypeptide chain structure of Nr-CAM reported previously, we show that Bravo has a heterodimer structure composed of an alpha chain of M(r) 140/130 and a beta chain of 60-80 kD. As with L1 and Ng-CAM, the two chains of Bravo are generated from an intact polypeptide by cleavage at identical locations and conserved sites within all three molecules (Ser-Arg/Lys-Arg). The similar domain composition and heterodimer structure, as well as the 40% amino acid sequence identity of these molecules, defines them as an evolutionarily related subgroup of CAMs. The relationship of Bravo to molecules known to be involved in cell adhesion and process outgrowth, combined with its pattern of expression and numerous potential isoforms, suggests a complex role for this molecule in cell interactions during neural development.
神经系统中多种细胞表面分子在发育过程中确定细胞间相互作用方面发挥着重要作用。我们利用一种设计用于产生针对特定分子量神经表面分子的单克隆抗体的方法,先前已报道了在发育中的鸟类视网膜顶盖系统中发现的表面蛋白Bravo。在视网膜中发育的视神经纤维上可通过免疫检测到Bravo,但在顶盖中相同纤维的远端区域则不存在。我们已分离出包含Bravo整个编码区的cDNA克隆,包括含有五个cDNA替代序列的克隆。这些推测的替代剪接序列编码长度从10至93个氨基酸不等的多肽片段,预计既有胞外区域也有胞内区域。推导的Bravo一级结构显示,与细胞粘附分子(CAMs)鸡Ng - CAM和小鼠L1一样,Bravo由六个免疫球蛋白样结构域、五个纤连蛋白III型重复序列、一个跨膜结构域和一个短的胞质区域组成。最近,报道了一种相关分子Nr - CAM的cDNA序列,并讨论了它与Bravo可能的同一性(格鲁梅特,M.,V. 毛罗,M. P. 伯贡,G. E. 埃德尔曼,和B. A. 坎宁安。1991. 《细胞生物学杂志》113:1399 - 1412)。在此我们证实了这种同一性,而且表明在发育中的视网膜中,Bravo存在于米勒胶质细胞的突起和终足上。与先前报道的Nr - CAM的单多肽链结构不同,我们表明Bravo具有异二聚体结构,由分子量为140/130的α链和60 - 80 kD的β链组成。与L1和Ng - CAM一样,Bravo的两条链是通过在所有三个分子内相同位置和保守位点(Ser - Arg/Lys - Arg)处的切割从完整多肽产生的。这些分子相似的结构域组成和异二聚体结构,以及40%的氨基酸序列同一性,将它们定义为CAMs的一个进化相关亚组。Bravo与已知参与细胞粘附和突起生长的分子之间的关系,连同其表达模式和众多潜在的异构体,表明该分子在神经发育过程中的细胞相互作用中具有复杂的作用。