Imperial College London
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DrMichaelHohl

Faculty of Medicine, Department of Infectious Disease

Research Associate
 
 
 
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Contact

 

m.hohl Website

 
 
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Location

 

Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Seeger:2012:10.1371/journal.pone.0037845,
author = {Seeger, MA and Mittal, A and Velamakanni, S and Hohl, M and Schauer, S and Salaa, I and Grütter, MG and van, Veen HW},
doi = {10.1371/journal.pone.0037845},
journal = {PLoS One},
title = {Tuning the drug efflux activity of an ABC transporter in vivo by in vitro selected DARPin binders.},
url = {http://dx.doi.org/10.1371/journal.pone.0037845},
volume = {7},
year = {2012}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - ABC transporters use the energy from binding and hydrolysis of ATP to import or extrude substrates across the membrane. Using ribosome display, we raised designed ankyrin repeat proteins (DARPins) against detergent solubilized LmrCD, a heterodimeric multidrug ABC exporter from Lactococcus lactis. Several target-specific DARPin binders were identified that bind to at least three distinct, partially overlapping epitopes on LmrD in detergent solution as well as in native membranes. Remarkably, functional screening of the LmrCD-specific DARPin pools in L. lactis revealed three homologous DARPins which, when generated in LmrCD-expressing cells, strongly activated LmrCD-mediated drug transport. As LmrCD expression in the cell membrane was unaltered upon the co-expression of activator DARPins, the activation is suggested to occur at the level of LmrCD activity. Consistent with this, purified activator DARPins were found to stimulate the ATPase activity of LmrCD in vitro when reconstituted in proteoliposomes. This study suggests that membrane transporters are tunable in vivo by in vitro selected binding proteins. Our approach could be of biopharmaceutical importance and might facilitate studies on molecular mechanisms of ABC transporters.
AU  - Seeger,MA
AU  - Mittal,A
AU  - Velamakanni,S
AU  - Hohl,M
AU  - Schauer,S
AU  - Salaa,I
AU  - Grütter,MG
AU  - van,Veen HW
DO  - 10.1371/journal.pone.0037845
PY  - 2012///
TI  - Tuning the drug efflux activity of an ABC transporter in vivo by in vitro selected DARPin binders.
T2  - PLoS One
UR  - http://dx.doi.org/10.1371/journal.pone.0037845
UR  - https://www.ncbi.nlm.nih.gov/pubmed/22675494
UR  - http://hdl.handle.net/10044/1/55306
VL  - 7
ER  -
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