Tojo S, Betchaku T, Ziccardi V J, Wyatt G R
J Cell Biol. 1978 Sep;78(3):823-38. doi: 10.1083/jcb.78.3.823.
Fat body cells of silkmoth pupae (Hyalophora cecropia ) contain granules, showing a less dense outer zone and a denser, often crystalline, inner portion appear after cocoon spinning and increase until the larval-pupal ecdysis; more granules are formed in females than in males. Urate granules, appearing fibrous in internal structure, first form about the same time, but their accumulation is more gradual, and continues in the pupa. Both types have been isolated by centrifugation. Protein granules dissolve in buffers to yield proteins 1 and 2, with distinct electrophoretic and antigenic properties. These proteins have been isolated individually from pupal fat body extracts by using their different thermal stabilities in phosphate buffer containing MgCl2 and (NH4)2SO4, respectively, and purification was completed by gel chromatography. Protein 1 has a molecular weight of 480,000 and a subunit of 85,000 daltons, while protein 2 gives values of 530,000 and 89,000, respectively. Their amino acid compositions are similar but distinct. Proteins 1 and 2 accumulate in the hemolymph, beginning 3 days before spinning, reach maximal levels at spinning, and then decline in the hemolymph while granules are formed in the fat body, although the total hemolymph protein concentration does not decline at this time. It is concluded that the fat body of the late, feeding larva synthesizes two related "storage proteins" and secretes them in partially crystalline granules as protein reserves for metamorphosis.
蚕蛾蛹(樗蚕蛾)的脂肪体细胞含有颗粒,这些颗粒在结茧后出现,外部区域密度较低,内部部分密度较高,通常呈结晶状,并且数量不断增加,直至幼虫 - 蛹蜕皮期;雌性形成的颗粒比雄性更多。尿酸盐颗粒的内部结构呈纤维状,大约在同一时间开始形成,但其积累过程较为缓慢,在蛹期仍持续积累。这两种颗粒都已通过离心分离出来。蛋白质颗粒可在缓冲液中溶解,产生具有不同电泳和抗原特性的蛋白质1和蛋白质2。分别利用它们在含有MgCl2和(NH4)2SO4的磷酸盐缓冲液中的不同热稳定性,从蛹脂肪体提取物中单独分离出这些蛋白质,并通过凝胶色谱法完成纯化。蛋白质1的分子量为480,000,亚基为85,000道尔顿,而蛋白质2的分子量分别为530,000和89,000。它们的氨基酸组成相似但又有所不同。蛋白质1和蛋白质2在血淋巴中积累,在结茧前3天开始,在结茧时达到最高水平,然后在血淋巴中含量下降,与此同时脂肪体中形成颗粒,尽管此时血淋巴总蛋白浓度并未下降。得出的结论是,晚期取食幼虫的脂肪体合成两种相关的“储存蛋白”,并将它们以部分结晶颗粒的形式分泌出来,作为变态过程中的蛋白质储备。