α-Synuclein and β-synuclein enhance secretion protein production in baculovirus expression vector system

被引：0
作者：
Chao-Yi Teng
Shou-Lin Chang
Meng-Feng Tsai
Tzong-Yuan Wu
机构：
[1] National Tsing Hua University,Institute of Bioinformatics and Structural Biology
[2] Da Yeh University,Department of Molecular Biotechnology
[3] Chung Yuan Christian University,Department of Bioscience Technology
[4] Chung Yuan Christian University,Center of Nanotechnology
来源：
Applied Microbiology and Biotechnology | 2013年 / 97卷
关键词：
Baculovirus; Calreticulin; α-Synuclein; β-Synuclein; Secreted proteins;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
The baculovirus expression vector system (BEVS) is widely used as a tool for expressing of recombinant proteins in insect cells or larvae. However, the expression level of secretion pathway proteins is often lower than that of cytosolic and nucleus proteins. Thus, we attempted to improve production of secreted proteins by using a secretory alkaline phosphatase-EGFP fusion protein (SEFP)-based bi-cistronic baculovirus vector to identify chaperones that have potential on boosting secreted protein production. As co-expressed SEFP with a chaperone, calreticulin (CALR), it was found that the secreted SEFP enzyme activity can be boosted up to twofold. This result demonstrated the SEFP-based bi-cistronic approach can be used to identify the genes that can enhance secretion protein production in BEVS. Thus, the chaperone activity of α-synuclein (α-syn) and β-synuclein (β-syn) was evaluated in cells co-expressed with SEFP and compared that with CALR by analyzing localization, alkaline phosphatase enzyme activity, and mRNA expression levels of SEFP. Our results showed that SEFP enzyme activity from cells co-expressed with both synuclein proteins can be enhanced up to 2.3-fold and this increment was better than that caused by CALR. Moreover, this enhancement might arise from the transcription enhancement or higher RNA stability. By this novel approach, we provided evidences that α- and β-syn can enhance secretion proteins production in BEVS.
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页码：3875 / 3884
页数：9
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