Perlecan domain V of Drosophila melanogaster -: Sequence, recombinant analysis and tissue expression
Perlecan domain V of Drosophila melanogaster -: Sequence, recombinant analysis and tissue expression
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DOI：
10.1046/j.1432-1327.2000.01337.x
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发表时间：
2000-06-01
期刊：
影响因子：
--
通讯作者：
Baumgartner, S
中科院分区：
文献类型：
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作者：
Friedrich, MVK;Schneider, M;Baumgartner, S
基膜蛋白多糖的C末端结构域V在细胞外基质和细胞间相互作用中起重要作用。果蝇的708个氨基酸残基的同源序列与小鼠Perlecan结构域V有33%的同源性，它由三个层粘连蛋白G型(LG)和表皮生长因子样(EG)模块组成，但缺乏EG3模块和哺乳动物Perlecans中的一个连接区。在哺乳动物细胞中重组果蝇Perlecan结构域V产生了一个100 kDa的蛋白，该蛋白被折叠成一个由三个球形LG结构域组成的线性阵列。与小鼠不同，果蝇的V结构域不被糖胺聚糖和内源性蛋白分解修饰，这是因为缺乏连接区。它对肝素和硫脂具有中等亲和力，但不与鸡α-肌营养不良蛋白或各种哺乳动物基底膜蛋白结合。果蝇V区LG3中的单个RGD序列也不能介导细胞黏附。用抗果蝇结构域V的抗体进行免疫印迹实验表明，果蝇细胞系可产生约450 kDa的蛋白多糖，在胚胎发育的不同阶段也有较强的表达，并定位于几个基底膜区。这表明，对于哺乳动物物种来说，Perlecan在果蝇发育过程中发挥着独特的作用。
The C-terminal domain V of the basement membrane proteoglycan perlecan was previously shown to play a major role in extracellular matrix and cell interactions. A homologous sequence of 708 amino-acid residues from Drosophila has now been shown to be 33% identical to mouse perlecan domain V. It consists of three laminin G-type (LG) and epidermal growth factor-like (EG) modules but lacks the EG3 module and a link region found in mammalian perlecans. Recombinant production of Drosophila perlecan domain V in mammalian cells yielded a 100-kDa protein which was folded into a linear array of three globular LG domains. Unlike the mouse counterpart, domain V from Drosophila was not modified by glycosaminoglycans and endogenous proteolysis, due to the absence of the link region. It showed moderate affinities for heparin and sulfatides but did not bind to chick alpha-dystroglycan or to various mammalian basement membrane proteins. A single RGD sequence in LG3 of Drosophila domain V was also incapable of mediating cell adhesion. Production of a proteoglycan form of perlecan (approximate to 450 kDa) in one Drosophila cell line could be demonstrated by immunoblotting with antibodies against Drosophila domain V. A strong expression was also found by in situ hybridization and immunohistology at various stages of embryonic development and expression was localized to several basement membrane zones. This indicates, as for mammalian species, a distinct role of perlecan during Drosophila development.