Expression and characterization of two functional vacuolar sorting receptor (VSR) proteins, BP-80 and AtVSR4 from culture media of transgenic tobacco BY-2 cells
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摘要Vacuolar sorting receptors (VSRs) are type I integral membrane family proteins that mediate protein transport from late Golgi or trans-Golgi network (TGN) to vacuole in plant cells. The N-terminus of VSR is believed to be important for cargo binding while its transmembrane domain (TMD) and cytoplasmic tail (CT) are essential for its correct subcellular localization. In this study, we have developed and tested an expression system using transgenic tobacco BY-2 cells to produce truncated VSR (VSRNT) proteins lacking the TMD/CT into the culture media. The expressed truncated VSRs (BP80NT and AtVSR4NT) are properly secreted into the culture media and bind specifically to the known vacuolar sorting determinants (VSDs) of various vacuolar proteins in a calcium-dependent manner in vitro. Therefore, since VSR cargo proteins are likely secreted into the culture media along with the truncated VSRs, potential applications of such expression system include identification of native VSR cargo proteins from the culture media of transgenic cells via LC-MS/MS analysis and large-scale purification of truncated VSR proteins for structural study to understand cargo-receptor interaction mechanisms in plants. (C) 2010 Elsevier Ireland Ltd. All rights reserved.
著者Suen PK, Shen JB, Sun SSM, Jiang LW
期刊名稱Plant Science
出版年份2010
月份7
日期1
卷號179
期次1-2
出版社ELSEVIER IRELAND LTD
頁次68 - 76
國際標準期刊號0168-9452
語言英式英語
關鍵詞BY-2 cells; Protein secretion; Receptor-ligand binding; Vacuolar sorting determinant; Vacuolar sorting receptor
Web of Science 學科類別Biochemistry & Molecular Biology; BIOCHEMISTRY & MOLECULAR BIOLOGY; Plant Sciences; PLANT SCIENCES

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