TY  - JOUR
A1  - Speck, Janina
A1  - Arndt, Katja Maren
A1  - Müller, Kristian M.
T1  - Efficient phage display of intracellularly folded proteins mediated by the TAT pathway
JF  - Protein engineering design & selection
N2  - Phage display with filamentous phages is widely applied and well developed, yet proteins requiring a cytoplasmic environment for correct folding still defy attempts at functional display. To extend applicability of phage display, we employed the twin-arginine translocation (TAT) pathway to incorporate proteins fused to the C-terminal domain of the geneIII protein into phage particles. We investigated functionality and display level of fluorescent proteins depending on the translocation pathway, which was the TAT, general secretory (SEC) or signal recognition particle (SRP) pathway mediated by the TorA, PelB or DsbA signal sequences, respectively. Importantly, for green fluorescent protein, yellow fluorescent protein and cyan fluorescent protein, only TAT, but not SEC or SRP, translocation led to fluorescence of purified phage particles, although all three proteins could be displayed regardless of the translocation pathway. In contrast, the monomeric red fluorescent protein mCherry was functionally displayed regardless of the translocation pathway. Hence, correct folding and fluorophor formation of mCherry is not limited to the cytosol. Furthermore, we successfully displayed firefly luciferase as well as an 83 kDa argonaute protein, both containing free cysteines. This demonstrates broad applicability of the TAT-mediated phagemid system for the display of proteins requiring cytoplasmic factors for correct folding and should prove useful for the display of proteins requiring incorporation of co-factors or oligomerization to gain function.
KW  - g3p
KW  - phagemid display
KW  - protein design
KW  - protein engineering
KW  - selection
Y1  - 2011
U6  - https://doi.org/10.1093/protein/gzr001
SN  - 1741-0126
VL  - 24
IS  - 6
SP  - 473
EP  - 484
PB  - Oxford Univ. Press
CY  - Oxford
ER  - 
TY  - CHAP
A1  - Kükenshöner, Tim
A1  - Jean-Christoph, N.
A1  - Speck, J.
A1  - Müller, Kristian M.
A1  - Arndt, Katja Maren
T1  - Targeting the microphthalmia associated transcription factor coiled coil domain with interfering peptides
T2  - The FEBS journal
Y1  - 2011
SN  - 1742-464X
VL  - 278
IS  - 6
SP  - 159
EP  - 159
PB  - Wiley-Blackwell
CY  - Malden
ER  -