锌离子转运体8(ZnT8)主要表达在β细胞的胰岛素分泌囊泡膜上，介导Zn2+在胰岛素分泌囊泡中聚集，与胰岛素的储存运输相关。我们前期实验发现不同基因型ZnT8功能有差异，ZnT8随着胰岛素的分泌能运输到细胞膜上，但差异和运输的机制尚不清楚。本课题将首先表达纯化出全长ZnT8蛋白，利用异源表达和脂质体蛋白重构系统、停流光谱法等技术研究基因型差异导致功能差异的机制，随后使用稳定转染、细胞分选、激光共聚焦等手段研究ZnT8膜运输与胰岛素分泌调控的关系，并制备ZnT8膜外表位特异性抗体，研究ZnT8膜运输作为功能性β细胞表面标志参与胰岛素分泌调控的机制。本课题将为功能性β细胞的鉴定提供一个可能的标志物，且胰岛素分泌调控提供一种新的机制。

Zinc transport 8 (ZnT8) is mainly located in β cell insulin secretory granules membrane and acts as a transport t enrich zinc ions required for normal processing and packing of insulin. Our previous study showed the functional differences among different genotypic ZnT8. And ZnT8 can traffic to cell membrane with insulin secretion. But the mechanism is still unclear. The project is focus on expression and purification the full length ZnT8. Then we will investigate the function difference mechanism using the heterologous cell expression, protein liposomal reconfigurable system, stopped-flow spectroscopy. It will explain the relation of genotype and phenotype. Meanwhile, we will research the relationship between ZnT8 membrane traffic and insulin secretion regulation using stably cell lines, cell sorting and confocal microscope. The specific antibody which target on the out-membrane domain of full length ZnT8 will be prepared. ZnT8 could be a functional marker on β cell and the mechanism of ZnT8 membrane traffic involved in insulin secretion regulation should be explained. The subject will identify a possible marker for functional β cells and provide a new mechanism for insulin secretion regulation.