Wyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, Boston, MA, 02115, USA.
Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Boston, MA, 02115, USA.
Nat Commun. 2018 Apr 3;9(1):1311. doi: 10.1038/s41467-018-03768-x.
Compartmentalization of proteins into organelles is a promising strategy for enhancing the productivity of engineered eukaryotic organisms. However, approaches that co-opt endogenous organelles may be limited by the potential for unwanted crosstalk and disruption of native metabolic functions. Here, we present the construction of synthetic non-endogenous organelles in the eukaryotic yeast Saccharomyces cerevisiae, based on the prokaryotic family of self-assembling proteins known as encapsulins. We establish that encapsulins self-assemble to form nanoscale compartments in yeast, and that heterologous proteins can be selectively targeted for compartmentalization. Housing destabilized proteins within encapsulin compartments afford protection against proteolytic degradation in vivo, while the interaction between split protein components is enhanced upon co-localization within the compartment interior. Furthermore, encapsulin compartments can support enzymatic catalysis, with substrate turnover observed for an encapsulated yeast enzyme. Encapsulin compartments therefore represent a modular platform, orthogonal to existing organelles, for programming synthetic compartmentalization in eukaryotes.
将蛋白质分隔到细胞器中是提高工程真核生物生产力的一种很有前途的策略。然而，内源性细胞器的共选方法可能受到不必要的串扰和对天然代谢功能的破坏的限制。在这里，我们提出了在真核酵母酿酒酵母中构建基于称为囊泡蛋白的原核自组装蛋白家族的合成非内源性细胞器。我们确定囊泡蛋白可以在酵母中自组装形成纳米级隔间，并且可以有选择地将异源蛋白靶向隔间化。将不稳定的蛋白质封装在囊泡蛋白隔间内可以提供体内抗蛋白水解降解的保护，而在隔间内部共定位时，分裂蛋白组分之间的相互作用得到增强。此外，囊泡蛋白隔间可以支持酶催化，观察到包封的酵母酶的底物周转率。因此，囊泡蛋白隔间代表了一个模块化平台，与现有细胞器正交，可用于在真核生物中编程合成隔间化。
