Lamy J, Compin S, Lamy J N
Arch Biochem Biophys. 1983 Jun;223(2):584-603. doi: 10.1016/0003-9861(83)90623-9.
Immunological cross-reactivities between isolated subunits of the scorpion Androctonus australis (Aa) and of the horseshoe crab Limulus polyphemus (Lp) hemocyanins were studied using subunit-specific antibodies prepared through immunoadsorption to pure immobilized subunits. Rocket immunoelectrophoreses of the various subunits of both hemocyanins were carried out at constant antigen concentration against the various subunit-specific antibody preparations. Then the data were analyzed through factorial correspondence analysis and compared to the respective intramolecular locations of the subunits in both hemocyanins. The results show that the dimeric subunits located in the central part of each (4 X 6)meric structure (Aa whole molecule and Lp half molecule) were strongly preserved. In addition, the (8 X 6)mer-forming subunit of Lp hemocyanin (LpIV) and the subunit occupying the same intramolecular position in Aa hemocyanin (Aa5A) were also strongly preserved. Besides the strong antigenic relatedness, less pronounced crossed immunoprecipitations or no precipitation at all were observed between subunits with homologous positions suggesting a minor structural and/or functional roles for these subunits. All the antigen-antibody combinations leading to an absence of immunoprecipitation were screened for the presence of soluble immunocomplexes by radioimmunological tests. In all cases, soluble immunocomplexes were observed. These results suggest the following evolution scenario. First, the central dimeric subunits, responsible of the dodecamer aggregation (Aa3C and 5B and LpV and VI) were already differentiated when Merostomata diverged from Arachnida. Second, the differentiation of the (8 X 6)mer-forming subunit occurred in the Merostomata ramification in a preserved subunit already possessing a functional advantage. Third, the differentiation of subunits Aa3A and Aa3B recently occurred in the scorpion ramification.
利用通过对纯固定化亚基进行免疫吸附制备的亚基特异性抗体，研究了蝎子澳大利亚杀人蝎（Aa）和鲎美洲鲎（Lp）血蓝蛋白分离亚基之间的免疫交叉反应性。针对各种亚基特异性抗体制剂，在恒定抗原浓度下对两种血蓝蛋白的各种亚基进行了火箭免疫电泳。然后通过因子对应分析对数据进行分析，并与两种血蓝蛋白中亚基各自的分子内位置进行比较。结果表明，位于每个（4×6）聚体结构（Aa全分子和Lp半分子）中心部分的二聚体亚基得到了强烈保留。此外，Lp血蓝蛋白的（8×6）聚体形成亚基（LpIV）和在Aa血蓝蛋白中占据相同分子内位置的亚基（Aa5A）也得到了强烈保留。除了强抗原相关性外，在同源位置的亚基之间观察到不太明显的交叉免疫沉淀或根本没有沉淀，这表明这些亚基的结构和/或功能作用较小。通过放射免疫测试筛选所有导致无免疫沉淀的抗原-抗体组合，以检测可溶性免疫复合物的存在。在所有情况下，均观察到可溶性免疫复合物。这些结果表明了以下进化情景。首先，当肢口纲动物从蛛形纲动物分化出来时，负责十二聚体聚集的中心二聚体亚基（Aa3C和5B以及LpV和VI）就已经分化。其次，（8×6）聚体形成亚基的分化发生在肢口纲动物分支中一个已经具有功能优势的保留亚基中。第三，亚基Aa3A和Aa3B的分化最近发生在蝎子分支中。
