8DAM

nbF3:nbE8:CaV beta subunit 1b complex

8DAM の概要

• エントリーDOI: 10.2210/pdb8dam/pdb
• 分子名称: Voltage-dependent L-type calcium channel subunit beta-1, nanobody F3, Nanobody E8, ... (6 entities in total)
• 機能のキーワード: nanobody-cacnb1b complex, transport protein, transport protein-immune system complex, transport protein/immune system
• 由来する生物種: Mus musculus (house mouse); 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 69621.50

構造登録者

Nirwan, N.,Minor, D.L. (登録日: 2022-06-13, 公開日: 2022-12-21, 最終更新日: 2024-10-30)

主引用文献

Morgenstern, T.J.,Nirwan, N.,Hernandez-Ochoa, E.O.,Bibollet, H.,Choudhury, P.,Laloudakis, Y.D.,Ben Johny, M.,Bannister, R.A.,Schneider, M.F.,Minor Jr., D.L.,Colecraft, H.M.
Selective posttranslational inhibition of Ca V beta 1 -associated voltage-dependent calcium channels with a functionalized nanobody.
Nat Commun, 13:7556-7556, 2022

PubMed Abstract: Ca influx through high-voltage-activated calcium channels (HVACCs) controls diverse cellular functions. A critical feature enabling a singular signal, Ca influx, to mediate disparate functions is diversity of HVACC pore-forming α and auxiliary Caβ-Caβ subunits. Selective Caα blockers have enabled deciphering their unique physiological roles. By contrast, the capacity to post-translationally inhibit HVACCs based on Caβ isoform is non-existent. Conventional gene knockout/shRNA approaches do not adequately address this deficit owing to subunit reshuffling and partially overlapping functions of Caβ isoforms. Here, we identify a nanobody (nb.E8) that selectively binds Caβ SH3 domain and inhibits Caβ-associated HVACCs by reducing channel surface density, decreasing open probability, and speeding inactivation. Functionalizing nb.E8 with Nedd4L HECT domain yielded Chisel-1 which eliminated current through Caβ-reconstituted Ca1/Ca2 and native Ca1.1 channels in skeletal muscle, strongly suppressed depolarization-evoked Ca influx and excitation-transcription coupling in hippocampal neurons, but was inert against Caβ-associated Ca1.2 in cardiomyocytes. The results introduce an original method for probing distinctive functions of ion channel auxiliary subunit isoforms, reveal additional dimensions of Caβ signaling in neurons, and describe a genetically-encoded HVACC inhibitor with unique properties.

PubMed: 36494348
DOI: 10.1038/s41467-022-35025-7

実験手法

X-RAY DIFFRACTION (2 Å)