Expression of a chloroplast ATP/ADP transporter in E. coli membranes: behind the Mistic strategy.

摘要: Eukaryotic membrane protein expression is still a major bottleneck for structural studies. Production in E. coli often leads to low level and/or aggregated proteins. In the last decade, strategies relying on new fusion revealed promising results. Fusion with amphipatic Mistic has been described favor membranes. Although, this approach already reported few proteins, little known about activity of fused We used strategy and obtained high levels chloroplast ATP/ADP transporter from A. thaliana (NTT1) characterized its transport properties. NTT1 very which can be recovered after vivo cleavage. Moreover, detailed molecular characterization purified mature form, or cleaved-off highlights correct fold latter one. Therefore, considering higher quantity form via approach, valuable obtaining quantities pure active proteins that are adequate