title: Α-synuclein and β-synuclein enhance secretion protein production in baculovirus expression vector system.
abstract: 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.
authors: Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan.
authors: Chao-Yi
authors: CY
authors: Teng
authors: Shou-Lin
authors: SL
authors: Chang
authors: Meng-Feng
authors: MF
authors: Tsai
authors: Tzong-Yuan
authors: TY
authors: Wu
mesh_terms: D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D037282:Calreticulin / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D051844:alpha-Synuclein; D051846:beta-Synuclein / Q000378:metabolism*
chemical_list: D037282:Calreticulin; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D051844:alpha-Synuclein; D051846:beta-Synuclein; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins