Marks N J, Shaw C, Maule A G, Davis J P, Halton D W, Verhaert P, Geary T G, Thompson D P
Comparative Neuroendocrinology Research Group, Queen's University of Belfast, Northern Ireland.
Biochem Biophys Res Commun. 1995 Dec 26;217(3):845-51. doi: 10.1006/bbrc.1995.2849.
Numerous FMRFamide-related peptides (FaRPs) have been isolated and sequenced from extracts of free-living and parasitic nematodes. The most abundant FaRP identified in ethanolic/methanolic extracts of the parasitic forms, Ascaris suum and Haemonchus contortus and from the free-living nematode, Panagrellus redivivus, was KHEYLRFamide (AF2). Analysis of the nucleotide sequences of cloned FaRP-precursor genes from C. elegans and, more recently, Caenorhabditis vulgaris identified a series of related FaRPs which did not include AF2. An acid-ethanol extract of Caenorhabditis elegans was screened radioimmunometrically for the presence of FaRPs using a C-terminally directed FaRP antiserum. Approximately 300 pmols of the most abundant immunoreactive peptide was purified to homogeneity and 30 pmols was subjected to Edman degradation analysis and gas-phase sequencing. The unequivocal primary structure of the heptapeptide, Lys-His-Glu-Tyr-Leu-Arg-Phe-NH2 (AF2) was determined following a single gas-phase sequencing run. The molecular mass of the peptide was determined using a time-of-flight mass spectrometer and was found to be 920 (MH+)+, which was consistent with the theoretical mass of C-terminally amidated AF2. These results indicate that C. elegans possesses more than one FaRP gene.
人们已经从自由生活和寄生线虫的提取物中分离并测序了许多与FMRF酰胺相关的肽（FaRPs）。在寄生线虫猪蛔虫和捻转血矛线虫以及自由生活线虫复苏短体线虫的乙醇/甲醇提取物中鉴定出的最丰富的FaRP是KHEYLRF酰胺（AF2）。对秀丽隐杆线虫以及最近的普通秀丽隐杆线虫克隆的FaRP前体基因的核苷酸序列分析，确定了一系列不包括AF2的相关FaRPs。使用C末端导向的FaRP抗血清，通过放射免疫测定法筛选秀丽隐杆线虫的酸乙醇提取物中是否存在FaRPs。将大约300 pmol最丰富的免疫反应性肽纯化至同质，30 pmol进行了埃德曼降解分析和气相测序。在单次气相测序运行后，确定了七肽Lys-His-Glu-Tyr-Leu-Arg-Phe-NH2（AF2）明确的一级结构。使用飞行时间质谱仪测定该肽的分子量，发现其为m/z 920（MH+），这与C末端酰胺化AF2的理论质量一致。这些结果表明秀丽隐杆线虫拥有不止一个FaRP基因。
