| First Author | Morgenstern TJ | Year | 2019 |
| Journal | Elife | Volume | 8 |
| PubMed ID | 31403402 | Mgi Jnum | J:278707 |
| Mgi Id | MGI:6357814 | Doi | 10.7554/eLife.49253 |
| Citation | Morgenstern TJ, et al. (2019) A potent voltage-gated calcium channel inhibitor engineered from a nanobody targeted to auxiliary CaVbeta subunits. Elife 8:e49253 |
| abstractText | Inhibiting high-voltage-activated calcium channels (HVACCs; CaV1/CaV2) 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 CaVbeta subunits. Nb.F3 by itself is functionally inert, providing a convenient vehicle to target active moieties to CaVbeta-associated channels. Nb.F3 fused to the catalytic HECT domain of Nedd4L (CaV-abetalator), an E3 ubiquitin ligase, ablated currents from diverse HVACCs reconstituted in HEK293 cells, and from endogenous CaV1/CaV2 channels in mammalian cardiomyocytes, dorsal root ganglion neurons, and pancreatic beta cells. In cardiomyocytes, CaV-abetalator redistributed CaV1.2 channels from dyads to Rab-7-positive late endosomes. This work introduces CaV-abetalator 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. |
