Quantification of PtdIns(3,4,5)P(3) dynamics in EGF-stimulated carcinoma cells: a comparison of PH-domain-mediated methods with immunological methods.

The Biochemical Journal
Shu-Chin YipJonathan M Backer

Abstract

Class IA PI3Ks (phosphoinositide 3-kinases) generate the secondary messenger PtdIns(3,4,5)P(3), which plays an important role in many cellular responses. The accumulation of PtdIns(3,4,5)P(3) in cell membranes is routinely measured using GFP (green fluorescent protein)-labelled PH (pleckstrin homology) domains. However, the kinetics of membrane PtdIns(3,4,5)P(3) synthesis and turnover as detected by PH domains have not been validated using an independent method. In the present study, we measured EGF (epidermal growth factor)-stimulated membrane PtdIns(3,4,5)P(3) production using a specific monoclonal anti-PtdIns(3,4,5)P(3) antibody, and compared the results with those obtained using PH-domain-dependent methods. Anti-PtdIns(3,4,5)P(3) staining rapidly accumulated at the leading edge of EGF-stimulated carcinoma cells. PtdIns(3,4,5)P(3) levels were maximal at 1 min, and returned to basal levels by 5 min. In contrast, membrane PtdIns(3,4,5)P(3) production, measured by the membrane translocation of an epitope-tagged (BTK)PH (PH domain of Bruton's tyrosine kinase), remained approx. 2-fold above basal level throughout 4-5 min of EGF stimulation. To determine the reason for this disparity, we measured the rate of PtdIns(3,4,5)P(3) hydr...Continue Reading

References

Nov 7, 1995·Proceedings of the National Academy of Sciences of the United States of America·M A LemmonJ Schlessinger
Mar 17, 1997·The EMBO Journal·R H ChenD Bar-Sagi
Aug 29, 1997·The Journal of Biological Chemistry·L E RamehL C Cantley
Nov 7, 1998·The Journal of Biological Chemistry·J M KavranM A Lemmon
Jun 29, 2000·Trends in Pharmacological Sciences·T BallaP Várnai
Feb 24, 2001·Ultramicroscopy·C RotschM Radmacher
Mar 26, 2002·FEBS Letters·Mark A LemmonCharles S Abrams
Nov 5, 2002·Journal of Cell Science·Vera DesMaraisSarah E Hitchcock-DeGregori
Jun 13, 2003·Molecular Biology of the Cell·Yi Elaine HuangPeter Devreotes
Jan 8, 2004·EMBO Reports·Marten PostmaPeter J M Van Haastert
Apr 30, 2005·The EMBO Journal·Jacco van RheenenKees Jalink
May 4, 2005·Trends in Cell Biology·C Peter DownesJohn M Lucocq
Feb 17, 2007·The Biochemical Journal·Severine I GharbiMichael D Waterfield

❮ Previous
Next ❯

Citations

Nov 17, 2009·Proceedings of the National Academy of Sciences of the United States of America·Haiyan WuJonathan M Backer
Dec 1, 2010·Proceedings of the National Academy of Sciences of the United States of America·Yong Ho BaePartha Roy
May 18, 2012·Molecular Biology of the Cell·Katsuhiro KatoKozo Kaibuchi
Jun 15, 2013·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Rachel Schnur Salamon, Jonathan M Backer
Jun 11, 2009·The Biochemical Journal·Gerald R V HammondRobin F Irvine
Mar 4, 2015·Biochimica Et Biophysica Acta·Gerald R V Hammond, Tamas Balla
Dec 22, 2016·The Journal of Cell Biology·Jennifer M DysonChristina A Mitchell
Dec 18, 2014·Nature·Emmanuel BoucrotHarvey T McMahon
Jun 28, 2016·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Qingshan YangVarsha Gandhi
Nov 14, 2019·Cell Reports·Annika BrosigBritta J Eickholt
May 18, 2021·Frontiers in Cell and Developmental Biology·Sarah E ConduitChristina A Mitchell
Jul 28, 2021·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Johanne Le CoqDaniel Lietha

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
Kevin D NiswenderDenis G Baskin
Nature Reviews. Molecular Cell Biology
Mark A Lemmon
© 2021 Meta ULC. All rights reserved