Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C.
Hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) of the plasma membrane by phospholipase C (PLC) generates two critical second messengers, inositol-1,4,5-trisphosphate and diacylglycerol. For the enzymatic reaction, PIP2 binds to positively charged amino acids in the pleckstrin homology do...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4676720?pdf=render |
| _version_ | 1811225191203733504 |
|---|---|
| author | Jong Bae Seo Seung-Ryoung Jung Weigang Huang Qisheng Zhang Duk-Su Koh |
| author_facet | Jong Bae Seo Seung-Ryoung Jung Weigang Huang Qisheng Zhang Duk-Su Koh |
| author_sort | Jong Bae Seo |
| collection | DOAJ |
| description | Hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) of the plasma membrane by phospholipase C (PLC) generates two critical second messengers, inositol-1,4,5-trisphosphate and diacylglycerol. For the enzymatic reaction, PIP2 binds to positively charged amino acids in the pleckstrin homology domain of PLC. Here we tested the hypothesis that positively charged divalent and multivalent cations accumulate around the negatively charged PIP2, a process called electrostatic charge shielding, and therefore inhibit electrostatic PIP2-PLC interaction. This charge shielding of PIP2 was measured quantitatively with an in vitro enzyme assay using WH-15, a PIP2 analog, and various recombinant PLC proteins (β1, γ1, and δ1). Reduction of PLC activity by divalent cations, polyamines, and neomycin was well described by a theoretical model considering accumulation of cations around PIP2 via their electrostatic interaction and chemical binding. Finally, the charge shielding of PIP2 was also observed in live cells. Perfusion of the cations into cells via patch clamp pipette reduced PIP2 hydrolysis by PLC as triggered by M1 muscarinic receptors with a potency order of Mg2+ < spermine4+ < neomycin6+. Accumulation of divalent cations into cells through divalent-permeable TRPM7 channel had the same effect. Altogether our results suggest that Mg2+ and polyamines modulate the activity of PLCs by controlling the amount of free PIP2 available for the enzymes and that highly charged biomolecules can be inactivated by counterions electrostatically. |
| first_indexed | 2024-04-12T09:02:39Z |
| format | Article |
| id | doaj.art-6957131b1439410bb6b999d2668420c6 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T09:02:39Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-6957131b1439410bb6b999d2668420c62022-12-22T03:39:11ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011012e014443210.1371/journal.pone.0144432Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C.Jong Bae SeoSeung-Ryoung JungWeigang HuangQisheng ZhangDuk-Su KohHydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) of the plasma membrane by phospholipase C (PLC) generates two critical second messengers, inositol-1,4,5-trisphosphate and diacylglycerol. For the enzymatic reaction, PIP2 binds to positively charged amino acids in the pleckstrin homology domain of PLC. Here we tested the hypothesis that positively charged divalent and multivalent cations accumulate around the negatively charged PIP2, a process called electrostatic charge shielding, and therefore inhibit electrostatic PIP2-PLC interaction. This charge shielding of PIP2 was measured quantitatively with an in vitro enzyme assay using WH-15, a PIP2 analog, and various recombinant PLC proteins (β1, γ1, and δ1). Reduction of PLC activity by divalent cations, polyamines, and neomycin was well described by a theoretical model considering accumulation of cations around PIP2 via their electrostatic interaction and chemical binding. Finally, the charge shielding of PIP2 was also observed in live cells. Perfusion of the cations into cells via patch clamp pipette reduced PIP2 hydrolysis by PLC as triggered by M1 muscarinic receptors with a potency order of Mg2+ < spermine4+ < neomycin6+. Accumulation of divalent cations into cells through divalent-permeable TRPM7 channel had the same effect. Altogether our results suggest that Mg2+ and polyamines modulate the activity of PLCs by controlling the amount of free PIP2 available for the enzymes and that highly charged biomolecules can be inactivated by counterions electrostatically.http://europepmc.org/articles/PMC4676720?pdf=render |
| spellingShingle | Jong Bae Seo Seung-Ryoung Jung Weigang Huang Qisheng Zhang Duk-Su Koh Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C. PLoS ONE |
| title | Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C. |
| title_full | Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C. |
| title_fullStr | Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C. |
| title_full_unstemmed | Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C. |
| title_short | Charge Shielding of PIP2 by Cations Regulates Enzyme Activity of Phospholipase C. |
| title_sort | charge shielding of pip2 by cations regulates enzyme activity of phospholipase c |
| url | http://europepmc.org/articles/PMC4676720?pdf=render |
| work_keys_str_mv | AT jongbaeseo chargeshieldingofpip2bycationsregulatesenzymeactivityofphospholipasec AT seungryoungjung chargeshieldingofpip2bycationsregulatesenzymeactivityofphospholipasec AT weiganghuang chargeshieldingofpip2bycationsregulatesenzymeactivityofphospholipasec AT qishengzhang chargeshieldingofpip2bycationsregulatesenzymeactivityofphospholipasec AT duksukoh chargeshieldingofpip2bycationsregulatesenzymeactivityofphospholipasec |