Secretion of annexin V from cultured cells requires a signal peptide.
Secretion of annexin V from cultured cells requires a signal peptide.
复制标题
DOI：
10.1053/plac.2001.0724
复制
发表时间：
2001-11
期刊：
影响因子：
3.8
通讯作者：
X. Wang;B. Campos;M. Kaetzel;J. Dedman
中科院分区：
文献类型：
--
作者：
X. Wang;B. Campos;M. Kaetzel;J. Dedman
膜联蛋白V是一种缺乏疏水信号肽的细胞内蛋白。然而，有几项研究报道膜联蛋白V的细胞外存在。在这项研究中，我们设计了膜联蛋白V的转基因，有或没有附加的分泌信号肽，并研究了转基因产物在COS-7细胞中的分泌。信号肽将膜联蛋白V靶向至转染细胞的内质网（ER）、高尔基体和培养基。相反，没有信号肽，膜联蛋白V只存在于细胞质中，在培养基中未检测到。为了证实我们的结果，我们还评估了两种培养细胞系中细胞外膜联蛋白V的存在：BeWo，胎盘滋养层绒毛膜癌细胞模型和人脐静脉内皮细胞（HUVEC）。我们的研究结果表明，膜联蛋白V的免疫定位在两种细胞的表面，但不能检测到在任何一种细胞类型的培养基。我们的研究结果表明，膜联蛋白V的分泌需要重组添加的疏水性信号肽和有限数量的内源性细胞表面膜联蛋白V BeWo和HUVEC细胞是最有可能来自相邻的受损细胞。
Annexin V is an intracellular protein that lacks a hydrophobic signal peptide. However, there are several studies reporting the extracellular presence of annexin V. In this study, we designed transgenes of annexin V with or without an attached secretory signal peptide and investigated the secretion of the transgene products in COS-7 cells. The signal peptide, targeted annexin V to the endoplasmic reticulum (ER), the Golgi and culture media of transfected cells. In contrast, without the signal peptide, annexin V was present only in the cytoplasm and was not detected in the medium. To confirm our results we also evaluated the presence of extracellular annexin V in two cultured cell lines: BeWo, a choriocarcinoma cell model of placental trophoblasts, and human umbilical vein endothelial cells (HUVEC). Our results showed that annexin V was immunolocalized on the surfaces of both cells but could not be detected in the culture medium of either cell type. Our results suggest that the secretion of annexin V required the recombinant addition of a hydrophobic signal peptide and that the limited quantities of endogenous cell surface annexin V on BeWo and HUVEC cells is most likely derived from adjacent damaged cells.