TY  - JOUR
A1  - Sachse, Rita
A1  - Wüstenhagen, Doreen Anja
A1  - Samalikova, Maria
A1  - Gerrits, Michael
A1  - Bier, Frank Fabian
A1  - Kubick, Stefan
T1  - Synthesis of membrane proteins in eukaryotic cell-free systems
JF  - Engineering in life sciences : Industry, Environment, Plant, Food
N2  - 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.
KW  - Cell-free protein expression
KW  - In vitro protein synthesis
KW  - Labeled membrane proteins
KW  - Synthetic glycoprotein
Y1  - 2013
U6  - https://doi.org/10.1002/elsc.201100235
SN  - 1618-0240
VL  - 13
IS  - 1
SP  - 39
EP  - 48
PB  - Wiley-Blackwell
CY  - Hoboken
ER  - 
TY  - JOUR
A1  - Zemella, Anne
A1  - Thoring, Lena
A1  - Hoffmeister, Christian
A1  - Samalikova, Maria
A1  - Ehren, Patricia
A1  - Wüstenhagen, Doreen Anja
A1  - Kubick, Stefan
T1  - Cell-free protein synthesis as a novel tool for directed glycoengineering of active erythropoietin
JF  - Scientific reports
N2  - As one of the most complex post-translational modification, glycosylation is widely involved in cell adhesion, cell proliferation and immune response. Nevertheless glycoproteins with an identical polypeptide backbone mostly differ in their glycosylation patterns. Due to this heterogeneity, the mapping of different glycosylation patterns to their associated function is nearly impossible. In the last years, glycoengineering tools including cell line engineering, chemoenzymatic remodeling and site-specific glycosylation have attracted increasing interest. The therapeutic hormone erythropoietin (EPO) has been investigated in particular by various groups to establish a production process resulting in a defined glycosylation pattern. However commercially available recombinant human EPO shows batch-to-batch variations in its glycoforms. Therefore we present an alternative method for the synthesis of active glycosylated EPO with an engineered O-glycosylation site by combining eukaryotic cell-free protein synthesis and site-directed incorporation of non-canonical amino acids with subsequent chemoselective modifications.
Y1  - 2018
U6  - https://doi.org/10.1038/s41598-018-26936-x
SN  - 2045-2322
VL  - 8
PB  - Nature Publ. Group
CY  - London
ER  -