Molecular properties and tissue distribution of 30K proteins as ommin-binding proteins from diapause eggs of the silkworm, Bombyx mori
Molecular properties and tissue distribution of 30K proteins as ommin-binding proteins from diapause eggs of the silkworm, Bombyx mori
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DOI：
10.1016/j.cbpb.2006.10.101
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发表时间：
2007-02-01
影响因子：
2.2
通讯作者：
Iino, Teruhiko
中科院分区：
文献类型：
--
作者：
Sawada, Hiroshi;Yamahama, Yumi;Iino, Teruhiko
我们曾报道从家蚕滞育卵的酸-甲醇提取物中纯化了一种ommin结合蛋白（OMBP），并与抗30 K蛋白抗血清发生反应。为了进一步阐明OMBP与30 K蛋白的关系，我们尝试通过双向聚丙烯酰胺凝胶电泳（2D-PAGE）对OMBP的N端氨基酸序列进行测定。我们观察到10个不同等电点的蛋白质斑点;斑点对应于30 kDa。根据其N端20个氨基酸的序列分析，这两个蛋白质分别属于家蚕血淋巴中的两种30 K蛋白（6 G1和19 G1）。这些蛋白质与伴刀豆球蛋白A和接骨木皮凝集素反应，因此预期会附着在多糖上。免疫组化结果表明，这些蛋白质主要分布在滞育卵的卵黄颗粒和浆膜中。这些结果表明，OMBP是由30 K蛋白质修饰的多糖。此外，通过RT-PCR分析，在滞育卵的早期胚胎阶段，观察到30 K蛋白mRNA的表达。30 K蛋白作为OMBP可能在浆膜形成过程中色氨酸代谢产物和色素的转运和积累中起重要作用。(c)2006爱思唯尔公司All rights reserved.
We previously reported the purification of an ommin-binding protein (OMBP) from an acid-methanol extract of diapause eggs of the silkworm and that OMBP reacted with the anti-30K proteins antiserum. In order to clarify the relationship between OMBP and the 30K proteins, we attempted to determine the sequence of the N-terminal amino acid of OMBP, which was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). We observed ten protein spots of various isoelectric points; the spots corresponded with 30 kDa. Based on the sequence of the N-terminal amino acid (20 residues), the spots belonged to two kinds of 30K proteins (6G1 and 19G1), which are known as the major plasma proteins in the larval hemolymph of the silkworm. The proteins are expected to attach to polysaccharide because they reacted with concanavalin A and elderberry bark lectin. Immumohistochemical observations clarified that the proteins were localized in yolk granules and serosa in the diapause egg. These results suggest that OMBP is composed of 30K proteins which were modified with polysaccharides. In addition, the expression of 30K proteins mRNA was observed at early embryonic stage in diapause eggs by RT-PCR analysis. The 30K proteins as OMBP may play an important role in the transport and accumulation of tryptophan metabolites and ommochrome during the formation of serosa. (c) 2006 Elsevier Inc. All rights reserved.