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首页> 外文期刊>Zoological Science >Preparation of two recombinant crustacean hyperglycemic hormones from the giant freshwater prawn, Macrobrachium rosenbergii, and their hyperglycemic activities.
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Preparation of two recombinant crustacean hyperglycemic hormones from the giant freshwater prawn, Macrobrachium rosenbergii, and their hyperglycemic activities.

机译：从巨型淡水虾罗氏沼虾制备两种重组甲壳类高血糖激素及其高血糖活性。

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摘要

Crustacean hyperglycemic hormone (CHH) is released from the X-organ/sinus gland complex located in the eyestalks, and regulates glucose levels in the hemolymph. In the giant freshwater prawn (Macrobrachium rosenbergii), two cDNAs encoding different CHH molecules were previously cloned by other workers. One of these (Mar-CHH-2) was expressed only in the eyestalks, whereas the other (Mar-CHH-L) was expressed in the heart, gills, antennal gland, and thoracic ganglion, but not in the eyestalks. However, their biological activities had not yet been characterized. Therefore, in this study, recombinant Mar-CHH-2 (rMar-CHH-2) and Mar-CHH-L (rMar-CHH-L) were produced using an E. coli expression system, by expression in bacterial cells and recovery in the insoluble fraction. Thereafter, rMar-CHH-2 and rMar-CHH-L were subjected to refolding and were subsequently purified by reversed-phase HPLC. The rMar-CHH-2 and rMar-CHH-L thus obtained exhibited the same disulfide bond arrangements as those of other CHHs reported previously, indicative of natural conformation. In in vivo bioassay, rMar-CHH-2 showed significant hyperglycemic activity, whereas rMar-CHH-L had no effect. These results indicate that Mar-CHH-L does not function as a CHH, but may have some other, unknown function..
机译：甲壳类高血糖激素（CHH）从位于眼球的X器官/窦腺复合物中释放，并调节血淋巴中的葡萄糖水平。在巨型淡水虾（罗氏沼虾）中，两个编码不同CHH分子的cDNA先前已被其他工人克隆。其中一种（Mar-CHH-2）仅在眼球中表达，而另一种（Mar-CHH-L）在心脏，腮，触角腺和胸神经节中表达，而在眼球中不表达。然而，它们的生物学活性尚未被表征。因此，在这项研究中，重组大肠杆菌Mar-CHH-2（rMar-CHH-2）和Mar-CHH-L（rMar-CHH-L）是使用大肠杆菌表达系统通过在细菌细胞中表达并在不溶部分。之后，对rMar-CHH-2和rMar-CHH-L进行重折叠，随后通过反相HPLC纯化。由此获得的rMar-CHH-2和rMar-CHH-L表现出与先前报道的其他CHH相同的二硫键排列，表明其天然构象。在体内生物测定中，rMar-CHH-2显示出明显的高血糖活性，而rMar-CHH-L没有作用。这些结果表明Mar-CHH-L不能用作CHH，但可能具有其他未知功能。

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