Conserved Neuron Promoting Activity in Drosophila and Vertebrate Laminin α1*
Conserved Neuron Promoting Activity in Drosophila and Vertebrate Laminin α1*
复制标题
DOI：
10.1074/jbc.271.30.18074
复制
发表时间：
1996-07
期刊：
影响因子：
--
通讯作者：
Y. Takagi;M. Nomizu;D. Gullberg;A. Mackrell;D. Keene;Yoshihiko Yamada;J. Fessler
中科院分区：
文献类型：
--
作者：
Y. Takagi;M. Nomizu;D. Gullberg;A. Mackrell;D. Keene;Yoshihiko Yamada;J. Fessler
用编码果蝇层粘连蛋白α链两个部分的构建体转染果蝇S2细胞。构建层粘连蛋白α III、I/II和G结构域编码的recαL。构建仅编码cooh最多12%的I/II结构域和G结构域的recαS。从培养基中分离出相应的多肽并对其进行了表征。recα -l链与内源性β和γ层粘连蛋白链部分形成二硫链异源三聚体。与正常果蝇层粘连蛋白一样，recαL或recαS的底物涂层支持原代果蝇胚胎细胞培养的神经元分化和神经突延伸。然而，在相同的低浓度下，在这些培养中，只有果蝇层粘连蛋白-1，而recαL和recαS都不支持肌发生。先前，一组重叠的十二肽覆盖了类似于recαS的鼠层粘连蛋白α1链的一个区域，并被合成并测试了细胞培养支持特性(Nomizu， M., Kim， W. H., Yamamura， K., Utani, a ., Otaka ., Roller， P. P., Kleinman， H. K., and Yamada， Y. (1995) J. Biol。化学，270,20583 -20590)。合成了6个最活跃的脊椎动物十二肽的果蝇层粘连蛋白α同源物，并作为果蝇胚胎细胞分化的底物进行了测试。含有果蝇序列SIKVGV或小鼠同源序列SIKVAV的多肽为神经突延伸提供了支持。
Drosophila S2 cells were transfected with constructs that code for two portions of the Drosophila laminin α chain. Construct recαL coded for domains III, I/II, and G of laminin α. Construct recαS coded for only the COOH-most 12% of the I/II domain and the G domain. The corresponding polypeptides were isolated and characterized from the culture media. The recαL chain partly formed disulfide-linked heterotrimers with the endogenously produced β and γ laminin chains. Like normal Drosophila laminin, a substrate coating of either recαL or recαS supported neuron differentiation and neurite extension of primary Drosophila embryo cell cultures. However, at the same low concentrations, only Drosophila laminin-1, but neither recαL nor recαS supported myogenesis in these cultures. Previously, an overlapping set of dodecapeptides that covered a region of the murine laminin α1 chain similar to recαS had been synthesized and tested for cell culture support properties (Nomizu, M., Kim, W. H., Yamamura, K., Utani, A., Otaka, A., Roller, P. P., Kleinman, H. K., and Yamada, Y. (1995) J. Biol. Chem. 270, 20583-20590). The Drosophila laminin α homologues of the six most active vertebrate dodecapeptides were now synthesized and tested as substrates for differentiation of primary Drosophila embryo cells. Peptides that contained either the Drosophila sequence SIKVGV or the murine homologue, SIKVAV, provided support for neurite extension.