Ollivaux Céline, Vinh Joëlle, Soyez Daniel, Toullec Jean-Yves
Université Pierre et Marie Curie-Paris 6, FRE CNRS 2852: Protéines: Biochimie Structurale et Fonctionnelle, Equipe Biogenèse des Peptides Isomères, Paris, France.
FEBS J. 2006 May;273(10):2151-60. doi: 10.1111/j.1742-4658.2006.05228.x.
Crustacean hyperglycemic hormone (CHH) and vitellogenesis-inhibiting hormone (VIH), produced by the X organ-sinus gland neurosecretory complex, belong to a peptide group referred to as the CHH family, which is widely distributed in arthropods. In this study, genetic variants and post-translationally modified isoforms of CHH and VIH were characterized in the European lobster Homarus gammarus. With the use of RP-HPLC and ELISA with specific antibodies that discriminate between stereoisomers of CHH and VIH, two groups of CHH-immunoreactive peaks were characterized from HPLC fractions of sinus gland extract (CHH A and CHH B); each group contained two variants (CHH and D-Phe3CHH). In the same way, two VIH-immunoreactive peaks (VIH and D-Trp4VIH) were demonstrated in HPLC fractions from sinus gland extract. The masses of these different neuropeptides were determined by FT-ICR MS: CHH A and CHH B spectra exhibited monoisotopic ions at 8557.05 Da and 8527.04 Da, respectively, and both VIH isomers displayed an m/z value of 9129.19 Da. Two full-length cDNAs encoding preprohomones of CHH A and CHH B and only one cDNA for VIH precursor were cloned and sequenced from X organ RNA. Comparison of CHH sequences between European lobster and other Astacoidea suggests that the most hydrophobic form appeared first during crustacean evolution.
由X器官 - 窦腺神经分泌复合体产生的甲壳动物高血糖激素(CHH)和卵黄发生抑制激素(VIH)属于一个被称为CHH家族的肽类群,该家族广泛分布于节肢动物中。在本研究中,对欧洲龙虾螯龙虾(Homarus gammarus)的CHH和VIH的基因变体及翻译后修饰的异构体进行了表征。利用RP - HPLC和能区分CHH和VIH立体异构体的特异性抗体进行ELISA,从窦腺提取物的HPLC馏分中鉴定出两组CHH免疫反应峰(CHH A和CHH B);每组包含两种变体(CHH和D - Phe3CHH)。同样,在窦腺提取物的HPLC馏分中证实了两个VIH免疫反应峰(VIH和D - Trp4VIH)。通过傅里叶变换离子回旋共振质谱(FT - ICR MS)测定了这些不同神经肽的质量:CHH A和CHH B的光谱分别在8557.05 Da和8527.04 Da处显示单同位素离子,两种VIH异构体的m/z值均为9129.19 Da。从X器官RNA中克隆并测序了编码CHH A和CHH B前体激素的两个全长cDNA以及仅一个VIH前体的cDNA。欧洲龙虾与其他螯虾科之间CHH序列的比较表明,在甲壳动物进化过程中,最疏水的形式最早出现。