IC-Tagging methodology applied to the expression of viral glycoproteins and the difficult-to-express membrane-bound IGRP autoantigen

被引:5
作者
Barreiro-Pineiro, Natalia [1 ]
Lostale-Seijo, Irene [1 ,2 ]
Varela-Calvino, Ruben [3 ]
Benavente, Javier [1 ]
Martinez-Costas, Jose M. [1 ]
机构
[1] Univ Santiago de Compostela, Dept Bioquim & Biol Mol, Grp Mol Virol, Ctr Singular Invest Quim Biol & Mat Mol CiQUS, Santiago De Compostela 15782, Spain
[2] Univ Santiago de Compostela, Ctr Singular Invest Quim Biol & Mat Mol CiQUS, Dept Quim Organ, Santiago De Compostela 15782, Spain
[3] Univ Santiago de Compostela, Fac Farm, Dept Bioquim & Biol Mol, Santiago De Compostela 15782, Spain
关键词
PROTEIN MU-NS; VIRUS FACTORIES; IDENTIFICATION; CELLS; ASSOCIATIONS; INCLUSIONS; INFECTION; VACCINES; MICE;
D O I
10.1038/s41598-018-34488-3
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We have previously developed a methodology to produce protein microspheres (MS) that can be loaded with proteins of interest in living cells through their C or N-terminal tagging with the so-called IC-Tag. The IC-Tagging method has many applications ranging from the production of immobilized enzymes for industrial use to the production of subunit vaccines due to its intrinsic adjuvancy. Here we show the adaptation of the IC-Tagging to work inside the endoplasmic reticulum and bacteria, allowing us to produce properly modified viral glycoproteins. Additionally, we were able to express the Islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP), whose expression remained elusive to date possibly due to its toxicity when over-expressed. IGRP is an antigen of enormous pharmaceutical interest as it is specifically targeted during the autoimmune response taking place in both the Non-Obese Diabetic (NOD) mice and type 1 diabetes (T1D) patients leading to the destruction of insulin-producing beta cells.
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页数:12
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