TY  - JOUR
A1  - Zimmermann, Iwan
A1  - Egloff, Pascal
A1  - Hutter, Cédric
A1  - Arnold, Fabian M.
A1  - Stohler, Peter
A1  - Bocquet, Nicolas
A1  - Hug, Melanie N.
A1  - Huber, Sylwia
A1  - Siegrist, Martin
A1  - Hetemann, Lisa
A1  - Gera, Jennifer
A1  - Gmür, Samira
A1  - Spies, Peter
A1  - Gygax, Daniel
A1  - Geertsma, Eric R.
A1  - Dawson, Roger
A1  - Seeger, Markus A.
T1  - Synthetic single domain antibodies for the conformational trapping of membrane proteins
T2  - eLife
N2  - Mechanistic and structural studies of membrane proteins require their stabilization in specific conformations. Single domain antibodies are potent reagents for this purpose, but their generation relies on immunizations, which impedes selections in the presence of ligands typically needed to populate defined conformational states. To overcome this key limitation, we developed an in vitro selection platform based on synthetic single domain antibodies named sybodies. To target the limited hydrophilic surfaces of membrane proteins, we designed three sybody libraries that exhibit different shapes and moderate hydrophobicity of the randomized surface. A robust binder selection cascade combining ribosome and phage display enabled the generation of conformation-selective, high affinity sybodies against an ABC transporter and two previously intractable human SLC transporters, GlyT1 and ENT1. The platform does not require access to animal facilities and builds exclusively on commercially available reagents, thus enabling every lab to rapidly generate binders against challenging membrane proteins.
KW  - Tools and ressources
KW  - Biochemistry and chemical biology
KW  - Structural biology and molecular biophysics
KW  - Membrane protein
KW  - Nanobody
KW  - Ribosome display
KW  - Phage display
KW  - In vitro selection
KW  - Conformational trapping
KW  - E. colo
Y1  - 2018
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/50355
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-503554
SN  - 2050-084X
N1  - Copyright Zimmermann et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
VL  - 7
IS  - e34317
SP  - 1
EP  - 32
PB  - eLife Sciences Publications
CY  - Cambridge
ER  -