家蚕贮藏蛋白Arylphorin结构解析及分子降解机制研究

Synthesis, absorption and utilization of storage protein, which has been known for its importance in insect growth and metamorphosis, has always been the focus in insect research. However, the research progress of storage protein is slow and mechanism of storage protein degradation is not fully elucidated. In this work, we will use structural biology tool to resolve the structure of silkworm storage protein in Arylphorin family. Afterwards according to their structural and protease cleavege information, we will design experiments to explore the mechanisms how storage protein is degradated, uptaken and utilized in silkworm. By use of protein purification, protein crystallization, crystal diffraction and data processing, we will get the high-resolution structure data of the silkworm storage protein. And then we will look for the specific degrading enzyme for silkworm storage protein with their structural information and silkworm microarray data. By analysis of expression characteristics and hydrolytic activity of silkworm protease, the degradation mechanisms of storage protein will be understood. Using small molecule inhibitors and RNAi method to the storage protein, we will confirm the relationship of silkworm storage protein and their degradation protease. This work will provide important reference for understanding development, metamorphosis and nutrient conversion process in silkworm.

贮藏蛋白合成、吸收和利用的研究一直是人们关注的焦点，其对于昆虫的生长和变态发育都有直接的影响。针对于目前贮藏蛋白研究进展缓慢，降解吸收机理尚未完全阐明的现状，本研究将利用生物科学的前沿手段- - 结构生物学的方法解析家蚕体内Arylphorin类贮藏蛋白的结构，利用获得蛋白质结构信息及相关蛋白酶切位点信息,设计实验探究家蚕贮藏蛋白在家蚕体内的降解和吸收利用机制。实验将通过蛋白纯化，蛋白结晶，晶体衍射，数据处理，结构解析等实验手段，获得高分辨率的家蚕贮藏蛋白晶体结构数据。利用蛋白质结构信息和家蚕基因组芯片表达相关数据，找到家蚕体内贮藏蛋白特异的降解酶，分析其在家蚕体内的表达特征和水解活性，并进一步利用小分子抑制剂和RNAi干涉实验对贮藏蛋白在家蚕体内被蛋白酶的降解过程进行研究，为人们认识理解家蚕贮藏蛋白的降解吸收机制及其在发育变态过程执行的功能提供参考。