Purification and characterization of an insulin-related peptide in the desert locust, Schistocerca gregaria:: immunolocalization, cDNA cloning, transcript profiling and interaction with neuroparsin
Purification and characterization of an insulin-related peptide in the desert locust, Schistocerca gregaria:: immunolocalization, cDNA cloning, transcript profiling and interaction with neuroparsin
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DOI：
10.1677/jme-07-0161
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发表时间：
2008-03-01
影响因子：
3.5
通讯作者：
Broeck, Jozef Vanden
中科院分区：
文献类型：
--
作者：
Badisco, Liesbeth;Claeys, Ilse;Broeck, Jozef Vanden
胰岛素超家族的成员不仅限于脊椎动物，而且在无脊椎动物物种中也被发现。在本报告中，我们提出了沙漠蝗虫体内的一种胰岛素相关肽SCG-胰岛素相关肽(IRP)的特性。该多肽是从心脏小体(CC)提取液中通过基于高效液相色谱(HPLC)的纯化策略分离得到的。随后的克隆和测序表明，编码的SCG-IRP前体显示了典型的胰岛素超家族成员的结构组织。此外，脑组织切片免疫细胞化学显示，SCG-IRP存在于大脑间部的正中神经分泌细胞中，并向CC的储存部投射。实时定量RT-PCR研究表明，SCG-IRP转录本存在于多种组织中，包括神经组织和脂肪体。此外，这些转录本在成年男性和女性的生殖周期中显示出组织和阶段依赖的时间调节。最后，我们证明了SCG-IRP在体外与重组神经帕斯汀相互作用，这是一种与脊椎动物IGF结合蛋白序列相似的蝗虫蛋白。
Members of the insulin superfamily are not restricted to vertebrates, but have also been identified in invertebrate species. In the current report, we present the characterization of Scg-insulin-related peptide (IRP), an insulin-related peptide in the desert locust, Schistocerca gregaria. This peptide was isolated from corpora cardiaca (CC) extracts by means of a high-performance liquid chromatography (HPLC)-based purification strategy. Subsequent cloning and sequencing of the corresponding cDNA revealed that the encoded Scg-IRP precursor displays the structural organization that is typical for members of the insulin superfamily. Moreover, immunocytochemistry on brain tissue sections demonstrated the presence of Scg-IRP in median neurosecretory cells of the pars intercerebralis and their projections towards the storage part of the CC. Quantitative real-time RT-PCR studies revealed the presence of Scg-IRP transcripts in a variety of tissues, including nervous tissue and fat body. Furthermore, these transcripts showed a tissue- and phase-dependent, temporal regulation during the reproductive cycle of adult males and females. Finally, we demonstrated that Scg-IRP interacts in vitro with a recombinant neuroparsin, a locust protein displaying sequence similarity with vertebrate IGF binding proteins.