Effects of ecdysone and juvenile hormone on the syntheis of specific proteins

研究了蜕皮激素和少年激素对家蚕幼虫和蛹凝集素样蛋白、蜕皮激素受体蛋白和促胸激素合成的影响。凝集素样蛋白出现在第4期的血淋巴中，并在第5期肠清洗后不久出现。该蛋白滴度的变化与蜕皮素滴度的变化几乎平行。这种蛋白具有很强的血凝素活性。利用该反应，分离并鉴定了该蛋白。分子量约350KD，被认为是88KD和90KD多肽的四聚体。糖，尤其是葡萄糖醛酸，能抑制血凝素的活性。从家蚕翅片中分离得到蜕皮激素受体蛋白。用放射性Ponasterone A测定其与表皮甾体的结合系数为4.1或4.3 nM。受体蛋白分子量分别为2.7KD和6.0KD。为了从家蚕基因组文库中提取受体基因，选择了一个类固醇激素的共同DNA结合域，合成了15个共同DNA序列。通过5龄幼虫研究了该蛋白在影像椎间盘中的含量变化。而结合活性/g总蛋白基本不变，有一定波动。研究了肉蝇滞育和非滞育培养中蜕皮素滴度的变化和影像盘的发育。在滞育幼虫和预蛹中，蜕皮激素含量在化蛹后上升，然后在化蛹前下降到几乎检测不到的水平。而非滞育幼虫在化蛹前数小时蜕皮素含量出现第二次升高。非滞育动物的象盘蛹发育较滞育动物早，但化蛹时间基本相同。

Effects of ecdysone and jevenile hormone on the synthesis of lectin-like protein, ecdysone receptor protein and prothoracicotropic hormone were studied in larvae and pupae of Bombyx silkworm. The lectin-like protein appeared in hemolymph just before 4th ecdysis and soon after the gut purge in the 5th instar. Change of titer of this protein is almost parallel to the change of ecdysone titer. This protein is of a strong hemagglutinin activity. Using this reaction, the protein was isolated and identified. Molecular weight is about 350KD, and it is thought as a tetramer of 88KD and 90KD polypeptide. Sugers, particularly, glucronic acid, inhibit hemagglutinin activity.Ecdysone receptor protein was isolated from the imaginal wing discs of Bombyx silkworm. Using radioactive Ponasterone A, binding coefficeient to ecdysteroid was determined as 4.1 or 4.3 nM. Molecular weight of the receptor protein was 2.7KD and 6.0KD. To fishing out of the receptor gene from genomic library of the silkworm, a common DNA binding domain of steroid hormones were selected and 15 sequences of common DNA sequence was synthesized. Change of amount of this protein in imaginal discs was studied through 5th instar larve. However, in terms of binding activity/g total protein was almost constant with some fluctiation.Change of ecdysone titer and development of imaginal discs were studied diapuse- and nondiapause-destined culture of the flesh fly. In diapause-destined larvae and prepupae, ecdysone content increased just after pupariation and then decreased to almost undetectable level before pupation. However, in nondiapause-destined larvae second increase of ecdysone content was observed several hours before pupation. Pupal development of imaginal discs was earlier in nondeapause-destined animals than deapause-destined ones, although pupation time was almost same between them.