Language
English
Publication Date
5-23-2025
Journal
Science Advances
DOI
10.1126/sciadv.adv1281
PMID
40408471
PMCID
PMC12101511
PubMedCentral® Posted Date
5-23-2025
PubMedCentral® Full Text Version
Post-print
Abstract
Action potentials are initiated and modulated at the axon initial segment (AIS) by highly clustered ion channels. Voltage-gated Kv1 potassium channels underlie most outward AIS K+ current. AIS Kv1 channels exist in a large protein complex including ADAM22, Caspr2, and LGI1. However, their clustering mechanisms remain unknown. Because Kv1 channels have a highly conserved PDZ-binding motif, we used CRISPR-based genome editing to screen 18 PDZ domain-containing proteins identified in our previous AIS proximity proteome for their AIS localization. Among these, we found that the scaffolding proteins SCRIB and PSD93 are highly enriched at the AIS. Using CRISPR-mediated knockout, cell surface clustering assays, and coimmunoprecipitation, we show that SCRIB and PSD93 bind to and are required for AIS Kv1 channel clustering, whereas SCRIB links the AIS Kv1 channel protein complex to the master AIS scaffolding protein AnkyrinG. These results define a hierarchy of scaffolding proteins that combine to cluster AIS Kv1 channels.
Keywords
PDZ Domains, Animals, Shaker Superfamily of Potassium Channels, Protein Binding, Humans, Axon Initial Segment, Rats, Membrane Proteins, HEK293 Cells, Axons, Kv Channel-Interacting Proteins
Published Open-Access
yes
Recommended Citation
Zhang, Wei; Palfini, Victoria L; Wu, Yu; et al., "A Hierarchy of Pdz Domain Scaffolding Proteins Clusters the Kv1 K+ Channel Protein Complex at the Axon Initial Segment" (2025). Faculty and Staff Publications. 5300.
https://digitalcommons.library.tmc.edu/baylor_docs/5300