One-shot design elevates functional expression levels of a voltage-gated potassium channel.
PROSS
membrane protein design
membrane protein expression
voltage‐gated potassium channels
Journal
Protein science : a publication of the Protein Society
ISSN: 1469-896X
Titre abrégé: Protein Sci
Pays: United States
ID NLM: 9211750
Informations de publication
Date de publication:
Jun 2024
Jun 2024
Historique:
revised:
03
04
2024
received:
27
09
2023
accepted:
05
04
2024
medline:
15
5
2024
pubmed:
15
5
2024
entrez:
15
5
2024
Statut:
ppublish
Résumé
Membrane proteins play critical physiological roles as receptors, channels, pumps, and transporters. Despite their importance, however, low expression levels often hamper the experimental characterization of membrane proteins. We present an automated and web-accessible design algorithm called mPROSS (https://mPROSS.weizmann.ac.il), which uses phylogenetic analysis and an atomistic potential, including an empirical lipophilicity scale, to improve native-state energy. As a stringent test, we apply mPROSS to the Kv1.2-Kv2.1 paddle chimera voltage-gated potassium channel. Four designs, encoding 9-26 mutations relative to the parental channel, were functional and maintained potassium-selective permeation and voltage dependence in Xenopus oocytes with up to 14-fold increase in whole-cell current densities. Additionally, single-channel recordings reveal no significant change in the channel-opening probability nor in unitary conductance, indicating that functional expression levels increase without impacting the activity profile of individual channels. Our results suggest that the expression levels of other dynamic channels and receptors may be enhanced through one-shot design calculations.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e4995Subventions
Organisme : Larry Marks Center for Brain Disorders
Organisme : Wilner Family Fund
Organisme : donation in memory of Sam Switzer
Organisme : Dr. Barry Sherman Institute for Medicinal Chemistry
Organisme : European Research Council through a Consolidator Award
ID : 815379
Organisme : Israel Science Foundation
ID : 1844
Organisme : Israel Science Foundation
ID : 349/22
Informations de copyright
© 2024 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.
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