多顺反子杆状病毒表达 SUGT1 可实现高产量的富含亮氨酸重复蛋白和蛋白复合物的重组表达。
Polycistronic baculovirus expression of SUGT1 enables high-yield production of recombinant leucine-rich repeat proteins and protein complexes.
机构信息
Protein Expression Laboratory, NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
Protein Expression Laboratory, NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
出版信息
Protein Expr Purif. 2022 May;193:106061. doi: 10.1016/j.pep.2022.106061. Epub 2022 Feb 4.
The SHOC2-MRAS-PPP1CA (SMP) complex is a holoenzyme that plays a vital role in the MAP kinase signaling pathway. Previous attempts to produce this challenging three-protein complex have relied on co-infection with multiple viruses and the use of affinity tags to attempt to isolate functional recombinant protein complexes. Leucine-rich repeat containing proteins have been historically challenging to express, and we hypothesized that co-expression of appropriate chaperones may be necessary for optimal production. We describe here how the SUGT1 chaperone can, in conjunction with polycistronic protein expression in baculovirus-infected insect cells, dramatically enhance production yield and quality of recombinant SHOC2, the SMP complex, and other leucine-rich repeat proteins.
SHOC2-MRAS-PPP1CA（SMP）复合物是一种全酶，在 MAP 激酶信号通路中起着至关重要的作用。以前生产这种具有挑战性的三蛋白复合物的尝试依赖于多种病毒的共感染和使用亲和标签来尝试分离功能性重组蛋白复合物。富含亮氨酸重复蛋白的表达历来具有挑战性，我们假设适当的伴侣蛋白的共表达可能是最佳生产所必需的。我们在这里描述了 SUGT1 伴侣蛋白如何与杆状病毒感染的昆虫细胞中的多顺反子蛋白表达相结合，显著提高重组 SHOC2、SMP 复合物和其他富含亮氨酸重复蛋白的产量和质量。
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