4.3 Article

Synthesis of membrane proteins in eukaryotic cell-free systems

Journal

ENGINEERING IN LIFE SCIENCES
Volume 13, Issue 1, Pages 39-48

Publisher

WILEY
DOI: 10.1002/elsc.201100235

Keywords

Cell-free protein expression; In vitro protein synthesis; Labeled membrane proteins; Synthetic glycoprotein

Funding

  1. German Ministry of Education and Research (BMBF) [0313965A, 0312039]

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Cell-free protein synthesis (CFPS) is a valuable method for the fast expression of difficult-to-express proteins as well as posttranslationally modified proteins. Since cell-free systems circumvent possible cytotoxic effects caused by protein overexpression in living cells, they significantly enlarge the scale and variety of proteins that can be characterized. We demonstrate the high potential of eukaryotic CFPS to express various types of membrane proteins covering a broad range of structurally and functionally diverse proteins. Our eukaryotic cell-free translation systems are capable to provide high molecular weight membrane proteins, fluorescent-labeled membrane proteins, as well as posttranslationally modified proteins for further downstream analysis.

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