Bhatt Manan, Di Iacovo Angela, Romanazzi Tiziana, Roseti Cristina, Cinquetti Raffaella, Bossi Elena
Laboratory of Cellular and Molecular Physiology, Department of Biotechnology and Life Sciences, University of Insubria, Via Dunant 3, 21100 Varese, Italy.
Experimental and Translational Medicine, University of Insubria, Via Ottorino Rossi 9, 21100 Varese, Italy.
Membranes (Basel). 2022 Sep 25;12(10):927. doi: 10.3390/membranes12100927.
After 50 years, the heterologous expression of proteins in oocytes is still essential in many research fields. New approaches and revised protocols, but also classical methods, such as the two-electrode voltage clamp, are applied in studying membrane transporters. New and old methods for investigating the activity and the expression of Solute Carriers (SLC) are reviewed, and the kinds of experiment that are still useful to perform with this kind of cell are reported. oocytes at the full-grown stage have a highly efficient biosynthetic apparatus that correctly targets functional proteins at the defined compartment. This small protein factory can produce, fold, and localize almost any kind of wild-type or recombinant protein; some tricks are required to obtain high expression and to verify the functionality. The methodologies examined here are mainly related to research in the field of membrane transporters. This work is certainly not exhaustive; it has been carried out to be helpful to researchers who want to quickly find suggestions and detailed indications when investigating the functionality and expression of the different members of the solute carrier families.
五十年来,卵母细胞中的蛋白质异源表达在许多研究领域仍然至关重要。新方法和修订后的方案,以及经典方法,如双电极电压钳,都被应用于膜转运蛋白的研究。本文综述了研究溶质载体(SLC)活性和表达的新旧方法,并报道了用这种细胞仍可进行的有用实验类型。完全成熟阶段的卵母细胞具有高效的生物合成装置,能够将功能性蛋白质正确靶向到特定的区室。这个小型蛋白质工厂几乎可以产生、折叠和定位任何种类的野生型或重组蛋白质;需要一些技巧来实现高表达并验证其功能。这里所研究的方法主要与膜转运蛋白领域的研究相关。这项工作当然并不详尽;开展此项工作是为了帮助那些在研究溶质载体家族不同成员的功能和表达时希望快速找到建议和详细指导的研究人员。