Tamashiro W K, Ehrenberg J P, Levy D A, Scott A L
Mol Biochem Parasitol. 1986 Mar;18(3):369-76. doi: 10.1016/0166-6851(86)90093-9.
Various methods of radioiodination were employed to identify peptides on the surface of Dirofilaria immitis microfilariae. Optimum surface radiolabelling occurred with the lactoperoxidase-catalyzed reaction. Two major peptides of 16 and 14 kDa were labelled by this method. These peptides were soluble in Nonidet P-40, were not glycosylated, and showed no signs of disulfide linkages. These peptides were immunoprecipitated by sera from D. immitis-infected dogs, but not by sera from uninfected dogs or sera from dogs with potentially cross-reactive nematode infections. Analysis of the 14 and 16 kDa peptides by two-dimensional gel electrophoresis revealed that the 16 kDa peptide was a single unit with a pI of 5.25 whereas the 14 kDa band was composed of three individual peptides with pI values ranging from 5.6 to 6.1. Iodination by chloramine T resulted in the same panel of labelled peptides but suffered from poor efficiency of 125I incorporation. The viability of microfilariae labelled by the standard Bolton-Hunter method decreased by 50% following the reaction which resulted in the labelling of a variety of internal components.
采用了多种放射性碘化方法来鉴定犬恶丝虫微丝蚴表面的肽段。通过乳过氧化物酶催化反应实现了最佳的表面放射性标记。用该方法标记出了两条主要的肽段，分子量分别为16 kDa和14 kDa。这些肽段可溶于去氧胆酸钠，无糖基化现象，也没有二硫键连接的迹象。这些肽段能被感染犬恶丝虫的犬血清免疫沉淀，但不能被未感染犬的血清或感染有潜在交叉反应性线虫的犬血清免疫沉淀。通过二维凝胶电泳分析14 kDa和16 kDa的肽段发现，16 kDa的肽段是一个单一单元，其等电点为5.25，而14 kDa的条带由三个单独的肽段组成，等电点值在5.6至6.1之间。用氯胺T进行碘化反应得到了相同的标记肽段图谱，但125I掺入效率较低。采用标准的博尔顿-亨特方法标记的微丝蚴在反应后活力下降了50%，该反应导致多种内部成分被标记。
