Improved Production of Class I Phosphatidylinositol 4,5-bisphosphate 3-kinase.

PI3K PIK3CA Phosphoinositide 3-kinase p110α p110β p110δ protein production p85α

Journal

Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496

Informations de publication

Date de publication:
20 Aug 2024
Historique:
received: 02 07 2024
revised: 05 08 2024
accepted: 18 08 2024
medline: 23 8 2024
pubmed: 23 8 2024
entrez: 22 8 2024
Statut: aheadofprint

Résumé

Phosphatidylinositol 4,5-bisphosphate 3-kinases (PI3K) are a family of kinases whose activity affects pathways needed for basic cell functions. As a result, PI3K is one of the most mutated genes in all human cancers and serves as an ideal therapeutic target for cancer treatment. Expanding on work done by other groups we improved protein yield to produce stable and pure protein using a variety of modifications including improved solubility tag, novel expression modalities, and optimized purification protocol and buffer. By these means, we achieved a 40-fold increase in yield for p110α/p85α and a 3-fold increase in p110α. We also used these protocols to produce comparable constructs of the β and δ isoforms of PI3K. Increased yield enhanced the efficiency of our downstream high throughput drug discovery efforts on the PIK3 family of kinases.

Identifiants

DOI: 10.1016/j.pep.2024.106582 PMID: 39173964
pubmed: 39173964
pii: S1046-5928(24)00154-2
doi: 10.1016/j.pep.2024.106582
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

106582

Informations de copyright

Copyright © 2024. Published by Elsevier Inc.

Déclaration de conflit d'intérêts

Declaration of Competing Interest The authors declare they have no conflicts of interest.

Auteurs

Simon Messing (S)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA. Electronic address: simon.messing@nih.gov.

Stephanie R T Widmeyer (SRT)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

John-Paul Denson (JP)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Jennifer Mehalko (J)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Vanessa E Wall (VE)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Matthew Drew (M)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Kelly Snead (K)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Min Hong (M)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Carissa Grose (C)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Dominic Esposito (D)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

William Gillette (W)

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.

Classifications MeSH