Details

Autor(en) / Beteiligte

• Morgenstern, Travis J
• Park, Jinseo
• Fan, Qing R
• Colecraft, Henry M

Titel

A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary Ca V β subunits

Ist Teil von

• eLife, 2019-08, Vol.8

Ort / Verlag

England

Erscheinungsjahr

2019

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• Inhibiting high-voltage-activated calcium channels (HVACCs; Ca 1/Ca 2) is therapeutic for myriad cardiovascular and neurological diseases. For particular applications, genetically-encoded HVACC blockers may enable channel inhibition with greater tissue-specificity and versatility than is achievable with small molecules. Here, we engineered a genetically-encoded HVACC inhibitor by first isolating an immunized llama nanobody (nb.F3) that binds auxiliary HVACC Ca β subunits. Nb.F3 by itself is functionally inert, providing a convenient vehicle to target active moieties to Ca β-associated channels. Nb.F3 fused to the catalytic HECT domain of Nedd4L (Ca -aβlator), an E3 ubiquitin ligase, ablated currents from diverse HVACCs reconstituted in HEK293 cells, and from endogenous Ca 1/Ca 2 channels in mammalian cardiomyocytes, dorsal root ganglion neurons, and pancreatic β cells. In cardiomyocytes, Ca -aβlator redistributed Ca 1.2 channels from dyads to Rab-7-positive late endosomes. This work introduces Ca -aβlator as a potent genetically-encoded HVACC inhibitor, and describes a general approach that can be broadly adapted to generate versatile modulators for macro-molecular membrane protein complexes.