Kempter B, Markl J, Brenowitz M, Bonaventura C, Bonaventura J
Biol Chem Hoppe Seyler. 1985 Jan;366(1):77-86. doi: 10.1515/bchm3.1985.366.1.77.
The hemocyanins of the horseshoe crab Limulus polyphemus (48-mer), the tarantula Eurypelma californicum (24-mer), and the lycosid spider Cupiennius salei (dodecamer, hexamer) were dissociated into subunits, the subunits isolated and studied by two-dimensional immunoelectrophoresis for interspecific cross-reactivities. Among the subunits a to g of Eurypelma on the one side, and I to VI of Limulus on the other, a number of cross-reactions were obtained which agree with the topologic subunit positions in the published models of quaternary structure: a = II, b-c = V-VI, d = IV, e = I, f = IIIb, g = IIIa (IIa). However, cross-reactivity was only strong in the following combinations: a/II, d/IV, b-c/V-VI (the monomers of the two heterodimers could not be correlated individually). A rather weak cross-reaction was obtained in the case of e/I and g/IIIa (IIa); a cross-reaction between f and IIIb was almost undetectable. On the other hand, f/IV clearly cross-reacted, and so did e/IIIa (IIa), which apparently is not in agreement with the two models of quaternary structure. These unexpected relationships, however, indicate the possible phylogeny of the subunits. Antiserum against Cupiennius hemocyanin precipitated subunit f of Eurypelma and subunit IV of Limulus and, moreover, revealed common antigen determinants present on these subunits. Denaturation of hemocyanin subunits of the three species with 8M urea yielded a completely different immunological behavior in that in all intra- and interspecific combinations the reaction of immunological identity was obtained. The published models of quaternary structure and a possible subunit phylogeny of cheliceratan hemocyanins is discussed in view of the present results and the results of the preceding paper. [Markl, J. et al. (1984) Hoppe-Seyler's Z. Physiol. Chem. 365, 619-631.]
将鲎（美洲鲎，48聚体）、加利福尼亚食鸟蛛（24聚体）和栉足蛛（十二聚体、六聚体）的血蓝蛋白解离成亚基，分离这些亚基，并通过二维免疫电泳研究种间交叉反应性。在食鸟蛛的a至g亚基与鲎的I至VI亚基之间，获得了许多交叉反应，这些反应与已发表的四级结构模型中的拓扑亚基位置一致：a = II，b - c = V - VI，d = IV，e = I，f = IIIb，g = IIIa（IIa）。然而，交叉反应仅在以下组合中较强：a/II、d/IV、b - c/V - VI（两个异二聚体的单体无法单独关联）。e/I和g/IIIa（IIa）的交叉反应较弱；f和IIIb之间的交叉反应几乎无法检测到。另一方面，f/IV明显发生交叉反应，e/IIIa（IIa）也是如此，这显然与两种四级结构模型不一致。然而，这些意外的关系表明了亚基可能的系统发育。针对栉足蛛血蓝蛋白的抗血清沉淀了食鸟蛛的亚基f和鲎的亚基IV，此外，还揭示了这些亚基上存在的共同抗原决定簇。用8M尿素使这三种物种血蓝蛋白亚基变性，产生了完全不同的免疫行为，即在所有种内和种间组合中都获得了免疫同一性反应。结合本文结果和前文结果，讨论了已发表的螯肢动物血蓝蛋白四级结构模型和可能的亚基系统发育。[马克尔，J.等人（1984年）《霍佩 - 赛勒生理化学杂志》365卷，619 - 631页。]
