Schoenmakers Ludo L J, den Uijl Max J, Postma Jelle L, van den Akker Tim A P, Huck Wilhelm T S, Driessen Arnold J M
Physical-Organic Chemistry, Institute for Molecules and Materials, Radboud University, Nijmegen 6525AJ, The Netherlands.
Groningen Biomolecular Sciences and Biotechnology, Molecular Biotechnology, University of Groningen, Groningen 9747 AG, The Netherlands.
Synth Biol (Oxf). 2024 May 10;9(1):ysae007. doi: 10.1093/synbio/ysae007. eCollection 2024.
Giant unilamellar vesicles (GUVs) provide a powerful model compartment for synthetic cells. However, a key challenge is the incorporation of membrane proteins that allow for transport, energy transduction, compartment growth and division. Here, we have successfully incorporated the membrane protein complex SecYEG-the key bacterial translocase that is essential for the incorporation of newly synthesized membrane proteins-in GUVs. Our method consists of fusion of small unilamellar vesicles containing reconstituted SecYEG into GUVs, thereby forming SecGUVs. These are suitable for large-scale experiments while maintaining a high protein:lipid ratio. We demonstrate that incorporation of SecYEG into GUVs does not inhibit its translocation efficiency. Robust membrane protein functionalized proteo-GUVs are promising and flexible compartments for use in the formation and growth of synthetic cells.
巨型单层囊泡（GUVs）为合成细胞提供了一个强大的模型隔室。然而，一个关键挑战是整合能够实现运输、能量转换、隔室生长和分裂的膜蛋白。在此，我们已成功地将膜蛋白复合物SecYEG（这是关键的细菌转位酶，对新合成膜蛋白的整合至关重要）整合到GUVs中。我们的方法包括将含有重组SecYEG的小单层囊泡融合到GUVs中，从而形成SecGUVs。这些适用于大规模实验，同时保持高蛋白：脂质比。我们证明将SecYEG整合到GUVs中不会抑制其转位效率。强大的膜蛋白功能化蛋白脂质体是用于合成细胞形成和生长的有前景且灵活的隔室。
