alpha-Synuclein and beta-synuclein enhance secretion protein production in baculovirus expression vector system

作者：Teng Chao Yi; Chang Shou Lin; Tsai Meng Feng; Wu Tzong Yuan^*

来源：Applied Microbiology and Biotechnology, 2013, 97(9): 3875-3884.

DOI：10.1007/s00253-012-4679-7

• Baculovirus
• Calreticulin
• alpha-Synuclein
• beta-Synuclein
• Secreted proteins

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 alpha-synuclein (alpha-syn) and beta-synuclein (beta-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 alpha- and beta-syn can enhance secretion proteins production in BEVS.

• 出版日期2013-5
• 单位清华大学