Department of Medical Pharmacology, Institute of Biomedical Sciences, Tokushima University, Tokushima, Japan.
Department of Health and Nutrition, Faculty of Nursing and Nutrition, The University of Shimane, Shimane, Japan.
Islets. 2022 Jan 1;14(1):1-13. doi: 10.1080/19382014.2021.1982325. Epub 2021 Oct 12.
The aim of this study was to identify genes that are specifically expressed in pancreatic islet β-cells (hereafter referred to as β-cells). Large-scale complementary DNA-sequencing analysis was performed for 3,429 expressed sequence tags derived from murine MIN6 β-cells, through homology comparisons using the GenBank database. Three individual ESTs were found to code for protease serine S1 family member 53 (). mRNA is processed into both a short and long form, which encode 482 and 552 amino acids, respectively. Transient overexpression of myc-tagged Prss53 in COS-7 cells showed that Prss53 was strongly associated with the luminal surfaces of organellar membranes and that it underwent signal peptide cleavage and N-glycosylation. Immunoelectron microscopy and western blotting revealed that Prss53 localized to mitochondria in MIN6 cells. Short hairpin RNA-mediated knockdown resulted in downregulation and and upregulation. JC-1 staining revealed that the mitochondria were depolarized in -knockdown MIN6 cells; however, no change was observed in glucose-stimulated insulin secretion. Our results suggest that mitochondrial Prss53 expression plays an important role in maintaining the health of β-cells.
本研究旨在鉴定特异性表达于胰岛 β 细胞（以下简称 β 细胞）的基因。通过与 GenBank 数据库进行同源性比较，对源自鼠 MIN6 β 细胞的 3429 个表达序列标签进行了大规模的 cDNA 测序分析。发现 3 个 EST 分别编码蛋白酶丝氨酸 S1 家族成员 53()。mRNA 被加工成短型和长型，分别编码 482 和 552 个氨基酸。在 COS-7 细胞中转染 myc 标记的 Prss53 后，发现 Prss53 与细胞器膜的腔面强烈相关，并经历信号肽切割和 N 糖基化。免疫电子显微镜和 Western blot 显示 Prss53 在 MIN6 细胞中定位于线粒体。短发夹 RNA 介导的 敲低导致 下调和 上调。JC-1 染色显示 -敲低 MIN6 细胞中线粒体去极化；然而，在葡萄糖刺激的胰岛素分泌中没有观察到变化。我们的结果表明，线粒体 Prss53 的表达在维持 β 细胞的健康中起着重要作用。
