Structural architecture of the human NALCN channelosome

Kschonsak, Marc; Chua, Han Chow; Weidling, Claudia; Chakouri, Nourdine; Noland, Cameron L.; Schott, Katharina; Chang, Timothy; Tam, Christine; Patel, Nidhi; Arthur, Christopher P.; Leitner, Alexander; Ben-Johny, Manu; Ciferri, Claudio; Pless, Stephan A.; Payandeh, Jian

doi:10.1038/s41586-021-04313-5

Cited by 26 publications

(32 citation statements)

References 56 publications

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“…In the center of the modular complex, Vps11 and Vps18 align antiparallel through their elongated α-solenoids, establishing a large interface area of 1972 Å 2 (Figure 1E-G, Figure 2, Figure 2-figure supplement 4), which resulted in the highest resolution obtained within HOPS (Figure 1-figure supplement 3) and is comparable with protein interfaces in other complexes with similar structural elements as in HOPS (e.g. Kschonsak et al, 2022). Interestingly, AlphaFold predicts a long unstructured region within Vps11 (Q760 to D784), resulting in an upper and lower part of the subunit.…”

Section: Structure Of the Hops Complexmentioning

confidence: 74%

Structure of the hops tethering complex, a lysosomal membrane fusion machinery

Moeller¹,

Shvarev²,

Schoppe³

2023

Biophysical Journal

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Section: Structure Of the Hops Complexmentioning

confidence: 74%

Structure of the hops tethering complex, a lysosomal membrane fusion machinery

Moeller¹,

Shvarev²,

Schoppe³

2023

Biophysical Journal

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“…Notably, during the submission process of this manuscript, another group published a cryo-EM structure of the human NALCN-FAM155A-UNC79-UNC80 complex 28 . Their structures of NALCN-FAM155A subcomplex and UNC79-UNC80 heterodimer are highly similar to ours (Supplementary Fig.…”

Section: Discussionmentioning

confidence: 99%

“…8d). In addition to the over-expressed NALCN, FAM155A, UNC79, and UNC80 proteins, they identified an endogenous calmodulin protein that is bound on the C-terminal helix of NALCN 28 . We also observed extra densities at similar positions not only in our current cryo-EM maps of the quaternary complex but also in our previously published NALCN-FAM155A subcomplex 15 (Supplementary Fig.…”

Section: Discussionmentioning

confidence: 99%

Structure and mechanism of NALCN-FAM155A-UNC79-UNC80 channel complex

Chen

2022

Nat Commun

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NALCN channel mediates sodium leak currents and is important for maintaining proper resting membrane potential. NALCN and FAM155A form the core complex of the channel, the activity of which essentially depends on the presence of both UNC79 and UNC80, two auxiliary proteins. NALCN, FAM155A, UNC79, and UNC80 co-assemble into a large hetero-tetrameric channel complex. Genetic mutations of NALCN channel components lead to neurodevelopmental diseases. However, the structure and mechanism of the intact channel complex remain elusive. Here, we present the cryo-EM structure of the mammalian NALCN-FAM155A-UNC79-UNC80 quaternary complex. The structure shows that UNC79-UNC80 form a large piler-shaped heterodimer which was tethered to the intracellular side of the NALCN channel through tripartite interactions with the cytoplasmic loops of NALCN. Two interactions are essential for proper cell surface localization of NALCN. The other interaction relieves the self-inhibition of NALCN by pulling the auto-inhibitory CTD Interacting Helix (CIH) out of its binding site. Our work defines the structural mechanism of NALCN modulation by UNC79 and UNC80.

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“…CryoEM data were processed using a combination of the RELION 49 and cisTEM 50 software packages, similarly as described previously 51 and as illustrated in Suppl. Figure 2 and 3.…”

Section: Cryoem Data Processingmentioning

confidence: 99%

CryoEM reveals unprecedented binding site for Na_V1.7 inhibitors enabling rational design of potent hybrid inhibitors

Kschonsak

Jao

Arthur

et al. 2022

Preprint

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The voltage-gated sodium (NaV) channel NaV1.7 has been identified as a potential novel pain target due to its striking human genetics. However, clinically available drugs (e.g. lidocaine, carbamazepine, etc.) are not selective among the nine NaV channel subtypes, NaV1.1-NaV1.9, and the two currently known classes of NaV1.7 subtype-selective inhibitors (aryl- and acylsulfonamides) have undesirable characteristics that may limit their development. Moreover, understanding of the structure-activity relationships of the acylsulfonamide class of NaV1.7 inhibitors, exemplified by the clinical development candidate GDC-0310, has been based solely on a single co-crystal structure of an arylsulfonamide inhibitor series. To advance inhibitor design targeting the NaV1.7 channel, we established an iterative system to routinely obtain high-resolution ligand bound NaV1.7 structures using cryogenic electron microscopy (cryo-EM). We report that GDC-0310 engages the NaV1.7 voltage-sensing domain 4 (VSD4) through an unexpected binding mode orthogonal to the arylsulfonamide class binding pose, which identifies a previously unknown ligand binding site in NaV channels. This finding enabled the design of a novel hybrid inhibitor series that bridges the aryl and acylsulfonamide binding pockets and allows for the generation of molecules with substantially differentiated structures and properties. Overall, this study highlights the power of cryo-EM methods to pursue challenging drug targets using iterative and high-resolution structure-guided inhibitor design. It also underscores an important role of the membrane bilayer in the discovery of selective NaV channel modulators.

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Structural architecture of the human NALCN channelosome

Cited by 26 publications

References 56 publications

Structure of the hops tethering complex, a lysosomal membrane fusion machinery

Structure of the hops tethering complex, a lysosomal membrane fusion machinery

Structure and mechanism of NALCN-FAM155A-UNC79-UNC80 channel complex

CryoEM reveals unprecedented binding site for Na_V1.7 inhibitors enabling rational design of potent hybrid inhibitors

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Structural architecture of the human NALCN channelosome

Cited by 26 publications

References 56 publications

Structure of the hops tethering complex, a lysosomal membrane fusion machinery

Structure of the hops tethering complex, a lysosomal membrane fusion machinery

Structure and mechanism of NALCN-FAM155A-UNC79-UNC80 channel complex

CryoEM reveals unprecedented binding site for NaV1.7 inhibitors enabling rational design of potent hybrid inhibitors

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Product

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CryoEM reveals unprecedented binding site for Na_V1.7 inhibitors enabling rational design of potent hybrid inhibitors