Sales M P, Pimenta P P, Paes N S, Grossi-de-Sá M F, Xavier-Filho J
Departamento de Bioquímica, Universidade Federal do Rio Grande do Norte, Natal, RN, Brasil.
Braz J Med Biol Res. 2001 Jan;34(1):27-34. doi: 10.1590/s0100-879x2001000100003.
The presence of chitin in midgut structures of Callosobruchus maculatus larvae was shown by chemical and immunocytochemical methods. Detection by Western blotting of cowpea (Vigna unguiculata) seed vicilins (7S storage proteins) bound to these structures suggested that C. maculatus-susceptible vicilins presented less staining when compared to C. maculatus-resistant vicilins. Storage proteins present in the microvilli in the larval midgut of the bruchid were recognized by immunolabeling of vicilins in the appropriate sections with immunogold conjugates. These labeling sites coincided with the sites labeled by an anti-chitin antibody. These results, taken together with those previously published showing that the lower rates of hydrolysis of variant vicilins from C. maculatus-resistant seeds by the insect's midgut proteinases and those showing that vicilins bind to chitin matrices, may explain the detrimental effects of variant vicilins on the development of C. maculatus larvae.
通过化学和免疫细胞化学方法证实了黄斑豆象幼虫中肠结构中几丁质的存在。通过蛋白质免疫印迹法检测与这些结构结合的豇豆(豇豆)种子伴刀豆球蛋白(7S储存蛋白)表明,与对黄斑豆象具有抗性的伴刀豆球蛋白相比,对黄斑豆象敏感的伴刀豆球蛋白染色较少。通过用免疫金缀合物对适当切片中的伴刀豆球蛋白进行免疫标记,识别出豆象幼虫中肠微绒毛中存在的储存蛋白。这些标记位点与抗几丁质抗体标记的位点一致。这些结果,连同先前发表的结果表明昆虫中肠蛋白酶对来自抗黄斑豆象种子的变体伴刀豆球蛋白的水解率较低,以及表明伴刀豆球蛋白与几丁质基质结合的结果,可能解释了变体伴刀豆球蛋白对黄斑豆象幼虫发育的有害影响。