The proteosomal degradation of fusion proteins cannot be predicted from the proteosome susceptibility of their individual components.
The proteosomal degradation of fusion proteins cannot be predicted from the proteosome susceptibility of their individual components.
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融合蛋白的蛋白酶体降解不能从其各个成分的蛋白酶体敏感性来预测。
DOI：
10.1110/ps.083443908
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Shneider,AlexanderM
中科院分区：
文献类型：
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作者：
Ilyinskii,PetrO;Meriin,AnatoliB;Gabai,VladimirL;Usachev,EvgenyV;Prilipov,AlexeiG;Thoidis,Galini;Shneider,AlexanderM
假设融合构建体的蛋白酶体加工特征可通过其组分的蛋白酶体敏感性总和进行预测。在本研究中，我们观察到由蛋白体可降解蛋白组成的融合构建体不一定会产生蛋白体可降解嵌合体。相反，蛋白酶体抗性蛋白的融合可能导致蛋白酶体可降解的复合物。我们先前证明，保守的流感蛋白可以统一为具有保护性的单一融合抗原，并且用蛋白体抗性和蛋白体可降解抗原的组合接种疫苗导致增强的T细胞应答。在本研究中，我们构建了保守流感蛋白NP、M1、NS 1和M2的蛋白体可降解突变体。然后将它们融合到不同位置的多分裂蛋白质中。这些融合构建体的稳定性和降解谱被证明取决于嵌合分子内各个蛋白质的相对位置。NP和M1或NS 1和M2的不稳定序列的组合导致快速蛋白体降解或蛋白体抗性二分融合突变体。然而，进一步将蛋白体可降解形式统一为单一的四部分融合分子导致相对稳定的嵌合蛋白。相反，向蛋白体抗性NP-M1-NS 1融合蛋白中添加蛋白体抗性野生型M2导致所得四部分多基因产物的稳定性降低，在一种情况下，其明显依赖于蛋白体。此外，M1的高度不稳定形式未能使野生型NP不稳定。总的来说，我们没有观察到任何导致蛋白体降解/非降解的多基因构建体的累加效应。
It is assumed that the proteosome‐processing characteristics of fusion constructs can be predicted from the sum of the proteosome sensitivity of their components. In the present study, we observed that a fusion construct consisting of proteosome‐degradable proteins does not necessarily result in a proteosome‐degradable chimera. Conversely, fusion of proteosome‐resistant proteins may result in a proteosome‐degradable composite. We previously demonstrated that conserved influenza proteins can be unified into a single fusion antigen that is protective, and that vaccination with combinations of proteosome‐resistant and proteosome‐degradable antigens resulted in an augmented T‐cell response. In the present study we constructed proteosome‐degradable mutants of conserved influenza proteins NP, M1, NS1, and M2. These were then fused into multipartite proteins in different positions. The stability and degradation profiles of these fusion constructs were demonstrated to depend on the relative position of the individual proteins within the chimeric molecule. Combining unstable sequences of either NP and M1 or NS1 and M2 resulted in either rapidly proteosome degraded or proteosome‐resistant bipartite fusion mutants. However, further unification of the proteosome‐degradable forms into a single four‐partite fusion molecule resulted in relatively stable chimeric proteins. Conversely, the addition of proteosome‐resistant wild‐type M2 to proteosome‐resistant NP–M1–NS1 fusion protein lead to the decreased stability of the resulting four‐partite multigene products, which in one case was clearly proteosome dependent. Additionally, a highly destabilized form of M1 failed to destabilize the wild‐type NP. Collectively, we did not observe any additive effect leading to proteosomal degradation/nondegradation of a multigene construct.