Fig. 1: Identification of functionally active nanobody binders for TREK-2. | Nature Communications

Fig. 1: Identification of functionally active nanobody binders for TREK-2.

From: Extracellular modulation of TREK-2 activity with nanobodies provides insight into the mechanisms of K2P channel regulation

Fig. 1

A Whole-cell currents recorded (at +40 mV) from oocytes expressing TREK-2 in the presence of a panel of nanobodies generated against TREK-2. Currents in the presence of extracellular nanobody (0.5 µM) were normalised to those in its absence. A threshold of 50% activation or inhibition was set in this screen (dotted lines). Tight binders within this panel were identified on the basis of producing a shift in size exclusion chromatography (SEC-shift) and shown in red. Asterisks mark the functionally active nanobodies plus an inactive tight binder chosen for further investigation. (Error bars represent mean ± S.D; n ≥ 5) B Crystal structures of the three functionally active nanobodies in complex with TREK-2. The nanobodies are shown in purple as surface representations whilst TREK-2 is shown in cartoon (green and gold). For CA10767 (Nb-Activator-67) the third ‘intracellular’ nanobody in the unit cell is shown in grey. A structure of the functionally inactive binder, CA10758 (Nb-Binder-58) is also shown in complex with TREK-2. In this case the nanobody binds as a dimer to the top of the Cap ___domain away from the extracellular TM/Pore loops that influence channel gating.

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