Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3

A. Toker, M. Meyer, K. K. Reddy, J R Falck, R. Aneja, S. Aneja, A. Parra, D. J. Burns, L. M. Ballas, L. C. Cantley

Research output: Contribution to journalArticle

533 Citations (Scopus)

Abstract

The effect of phosphoinositides on the activity of protein kinase C (PKC) isotypes was investigated. PKC α, βI, βII, γ, δ, ε, η, and ζ were expressed in baculovirus-infected insect cells and purified by column chromatography. The calcium-activated PKC isotypes α, βI, βII, and γ were not significantly activated by any of the phosphoinositides investigated (phosphatidylinositol-4-phosphate (PtdIns-4-P), PtdIns-3-P, PtdIns-4,5-P2, PtdIns-3,4-P2, and PtdIns-3,4,5-P3) when added in the presence of concentrations of phosphatidylserine that give maximal stimulation. The calcium-insensitive PKC isotypes δ, ε, and η also showed little response to PtdIns-3-P, PtdIns-4-P, or PtdIns-4,5-P2 when these lipids were added in the presence of phosphatidylserine. In contrast, PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 caused a 5-15-fold stimulation of these enzymes compared with phosphatidylserine alone. 50% maximal stimulation of PKC ε by PtdIns-3,4,5- P3 occurred when this lipid was present at about 1% of the carrier PtdIns- 4,5-P2 (about 100 nM). These lipids had little effect on baculovirus- expressed PKC ζ, which was constitutively active. A short chain version of PtdIns-3,4,5-P3, dioctanoyl-PtdIns-3,4,5-P3, activated PKC δ, ε, and η in the absence of other lipids, whereas a short chain version of PtdIns-4,5- P2, dihexanoyl-PtdIns-4,5-P2, did not. Since PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 are nominally absent in unstimulated cells and appear within seconds to minutes of stimulation by various cell activators, these lipids could act as second messengers to activate PKC δ, ε, or η in vivo.

Original languageEnglish (US)
Pages (from-to)32358-32367
Number of pages10
JournalJournal of Biological Chemistry
Volume269
Issue number51
StatePublished - 1994

Fingerprint

Phosphatidylinositol Phosphates
Protein Kinase C
Phosphatidylinositol 4,5-Diphosphate
Chemical activation
Phosphatidylinositols
Phosphatidylserines
Lipids
Baculoviridae
Calcium
Column chromatography
phosphatidylinositol 3,4-diphosphate
phosphatidylinositol 3,4,5-triphosphate
Second Messenger Systems
Insects
Chromatography
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Toker, A., Meyer, M., Reddy, K. K., Falck, J. R., Aneja, R., Aneja, S., ... Cantley, L. C. (1994). Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3 Journal of Biological Chemistry, 269(51), 32358-32367.

Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3 . / Toker, A.; Meyer, M.; Reddy, K. K.; Falck, J R; Aneja, R.; Aneja, S.; Parra, A.; Burns, D. J.; Ballas, L. M.; Cantley, L. C.

In: Journal of Biological Chemistry, Vol. 269, No. 51, 1994, p. 32358-32367.

Research output: Contribution to journalArticle

Toker, A, Meyer, M, Reddy, KK, Falck, JR, Aneja, R, Aneja, S, Parra, A, Burns, DJ, Ballas, LM & Cantley, LC 1994, 'Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3 ', Journal of Biological Chemistry, vol. 269, no. 51, pp. 32358-32367.
Toker, A. ; Meyer, M. ; Reddy, K. K. ; Falck, J R ; Aneja, R. ; Aneja, S. ; Parra, A. ; Burns, D. J. ; Ballas, L. M. ; Cantley, L. C. / Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3 In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 51. pp. 32358-32367.
@article{ccaea33f312b4af486d84c2fd3270430,
title = "Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3",
abstract = "The effect of phosphoinositides on the activity of protein kinase C (PKC) isotypes was investigated. PKC α, βI, βII, γ, δ, ε, η, and ζ were expressed in baculovirus-infected insect cells and purified by column chromatography. The calcium-activated PKC isotypes α, βI, βII, and γ were not significantly activated by any of the phosphoinositides investigated (phosphatidylinositol-4-phosphate (PtdIns-4-P), PtdIns-3-P, PtdIns-4,5-P2, PtdIns-3,4-P2, and PtdIns-3,4,5-P3) when added in the presence of concentrations of phosphatidylserine that give maximal stimulation. The calcium-insensitive PKC isotypes δ, ε, and η also showed little response to PtdIns-3-P, PtdIns-4-P, or PtdIns-4,5-P2 when these lipids were added in the presence of phosphatidylserine. In contrast, PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 caused a 5-15-fold stimulation of these enzymes compared with phosphatidylserine alone. 50{\%} maximal stimulation of PKC ε by PtdIns-3,4,5- P3 occurred when this lipid was present at about 1{\%} of the carrier PtdIns- 4,5-P2 (about 100 nM). These lipids had little effect on baculovirus- expressed PKC ζ, which was constitutively active. A short chain version of PtdIns-3,4,5-P3, dioctanoyl-PtdIns-3,4,5-P3, activated PKC δ, ε, and η in the absence of other lipids, whereas a short chain version of PtdIns-4,5- P2, dihexanoyl-PtdIns-4,5-P2, did not. Since PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 are nominally absent in unstimulated cells and appear within seconds to minutes of stimulation by various cell activators, these lipids could act as second messengers to activate PKC δ, ε, or η in vivo.",
author = "A. Toker and M. Meyer and Reddy, {K. K.} and Falck, {J R} and R. Aneja and S. Aneja and A. Parra and Burns, {D. J.} and Ballas, {L. M.} and Cantley, {L. C.}",
year = "1994",
language = "English (US)",
volume = "269",
pages = "32358--32367",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "51",

}

TY - JOUR

T1 - Activation of protein kinase C family members by the novel polyphosphoinositides PtdIns-3,4-P2 and PtdIns-3,4,5-P3

AU - Toker, A.

AU - Meyer, M.

AU - Reddy, K. K.

AU - Falck, J R

AU - Aneja, R.

AU - Aneja, S.

AU - Parra, A.

AU - Burns, D. J.

AU - Ballas, L. M.

AU - Cantley, L. C.

PY - 1994

Y1 - 1994

N2 - The effect of phosphoinositides on the activity of protein kinase C (PKC) isotypes was investigated. PKC α, βI, βII, γ, δ, ε, η, and ζ were expressed in baculovirus-infected insect cells and purified by column chromatography. The calcium-activated PKC isotypes α, βI, βII, and γ were not significantly activated by any of the phosphoinositides investigated (phosphatidylinositol-4-phosphate (PtdIns-4-P), PtdIns-3-P, PtdIns-4,5-P2, PtdIns-3,4-P2, and PtdIns-3,4,5-P3) when added in the presence of concentrations of phosphatidylserine that give maximal stimulation. The calcium-insensitive PKC isotypes δ, ε, and η also showed little response to PtdIns-3-P, PtdIns-4-P, or PtdIns-4,5-P2 when these lipids were added in the presence of phosphatidylserine. In contrast, PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 caused a 5-15-fold stimulation of these enzymes compared with phosphatidylserine alone. 50% maximal stimulation of PKC ε by PtdIns-3,4,5- P3 occurred when this lipid was present at about 1% of the carrier PtdIns- 4,5-P2 (about 100 nM). These lipids had little effect on baculovirus- expressed PKC ζ, which was constitutively active. A short chain version of PtdIns-3,4,5-P3, dioctanoyl-PtdIns-3,4,5-P3, activated PKC δ, ε, and η in the absence of other lipids, whereas a short chain version of PtdIns-4,5- P2, dihexanoyl-PtdIns-4,5-P2, did not. Since PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 are nominally absent in unstimulated cells and appear within seconds to minutes of stimulation by various cell activators, these lipids could act as second messengers to activate PKC δ, ε, or η in vivo.

AB - The effect of phosphoinositides on the activity of protein kinase C (PKC) isotypes was investigated. PKC α, βI, βII, γ, δ, ε, η, and ζ were expressed in baculovirus-infected insect cells and purified by column chromatography. The calcium-activated PKC isotypes α, βI, βII, and γ were not significantly activated by any of the phosphoinositides investigated (phosphatidylinositol-4-phosphate (PtdIns-4-P), PtdIns-3-P, PtdIns-4,5-P2, PtdIns-3,4-P2, and PtdIns-3,4,5-P3) when added in the presence of concentrations of phosphatidylserine that give maximal stimulation. The calcium-insensitive PKC isotypes δ, ε, and η also showed little response to PtdIns-3-P, PtdIns-4-P, or PtdIns-4,5-P2 when these lipids were added in the presence of phosphatidylserine. In contrast, PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 caused a 5-15-fold stimulation of these enzymes compared with phosphatidylserine alone. 50% maximal stimulation of PKC ε by PtdIns-3,4,5- P3 occurred when this lipid was present at about 1% of the carrier PtdIns- 4,5-P2 (about 100 nM). These lipids had little effect on baculovirus- expressed PKC ζ, which was constitutively active. A short chain version of PtdIns-3,4,5-P3, dioctanoyl-PtdIns-3,4,5-P3, activated PKC δ, ε, and η in the absence of other lipids, whereas a short chain version of PtdIns-4,5- P2, dihexanoyl-PtdIns-4,5-P2, did not. Since PtdIns-3,4-P2 and PtdIns- 3,4,5-P3 are nominally absent in unstimulated cells and appear within seconds to minutes of stimulation by various cell activators, these lipids could act as second messengers to activate PKC δ, ε, or η in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0028560447&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028560447&partnerID=8YFLogxK

M3 - Article

C2 - 7798235

AN - SCOPUS:0028560447

VL - 269

SP - 32358

EP - 32367

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 51

ER -