Production and purification of the PEAK pseudokinases for structural and functional studies.

Dimerization Oligomerization PEAK pseudokinases Protein expression Protein purification Scaffolds

Journal

Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271

Informations de publication

Date de publication:
2022
Historique:
entrez: 7 5 2022
pubmed: 8 5 2022
medline: 11 5 2022
Statut: ppublish

Résumé

The PEAK family of pseudokinases, which comprises PEAK1, PEAK2 and PEAK3, are newly identified scaffolds that dynamically assemble oncogenic signaling pathways known to contribute to the development of several aggressive cancers. A striking feature of this unique family of pseudokinase scaffolds is their large multi-domain structure, which allows them to achieve protein complex assemblies through their structural plasticity and functional versatility. Recent structural advances have begun to reveal the critical regulatory elements that control their function. Specifically, the dimer-dependent scaffolding activity of PEAK pseudokinases is emerging as a critical mechanism for their signaling function, in addition to their ability to hetero-associate to form higher-order regulatory networks to diversify and amplify their signaling output. Here, we present a suite of techniques that enable the efficient expression and purification of PEAK proteins for functional characterization.

Identifiants

pubmed: 35525538
pii: S0076-6879(22)00113-6
doi: 10.1016/bs.mie.2022.03.022
pii:
doi:

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1-35

Informations de copyright

Copyright © 2022 Elsevier Inc. All rights reserved.

Auteurs

Onisha Patel (O)

The Walter and Eliza Hall Institute of Medical Research, Parkville, Vic, Australia; Department of Medical Biology, University of Melbourne, Parkville, Vic, Australia. Electronic address: patel.o@wehi.edu.au.

Minglyanna Surudoi (M)

The Walter and Eliza Hall Institute of Medical Research, Parkville, Vic, Australia; Department of Medical Biology, University of Melbourne, Parkville, Vic, Australia.

Weiwen Dai (W)

The Walter and Eliza Hall Institute of Medical Research, Parkville, Vic, Australia; Department of Medical Biology, University of Melbourne, Parkville, Vic, Australia.

Joshua M Hardy (JM)

The Walter and Eliza Hall Institute of Medical Research, Parkville, Vic, Australia; Department of Medical Biology, University of Melbourne, Parkville, Vic, Australia.

Michael J Roy (MJ)

The Walter and Eliza Hall Institute of Medical Research, Parkville, Vic, Australia; Department of Medical Biology, University of Melbourne, Parkville, Vic, Australia.

Isabelle S Lucet (IS)

The Walter and Eliza Hall Institute of Medical Research, Parkville, Vic, Australia; Department of Medical Biology, University of Melbourne, Parkville, Vic, Australia. Electronic address: lucet.i@wehi.edu.au.

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