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Binding of PLCdelta1PH-GFP to PtdIns(4,5)P2 prevents inhibition of phospholipase C-mediated hydrolysis of PtdIns(4,5)P2 by neomycin.
Acta Pharmacol Sin. 2005 Dec; 26(12):1485-91.AP

Abstract

AIM

To investigate the effects of the pleckstrin homology (PH) domain of phospholipase C(delta1) (PLC(delta1)PH) on inhibition of phospholipase C (PLC)-mediated hydrolysis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] by neomycin.

METHODS

A fusion construct of green fluorescent protein (GFP) and PLC (delta1)PH (PLC(delta1)PH-GFP), which is known to bind PtdIns(4,5)P2 specifically, together with laser-scanning confocal microscopy, was used to trace PtdIns(4,5)P2 translocation.

RESULTS

Stimulation of the type 1 muscarinic receptor and the bradykinin 2 receptor induced a reversible PLC(delta1)PH-GFP translocation from the membrane to the cytosol in COS-7 cells. PLC inhibitor U73122 blocked the translocation. Wortmannin, a known PtdIns kinase inhibitor, did not affect the translocation induced by ACh, but blocked recovery after translocation, indicating that PtdIns(4,5)P2 hydrolysis occurs through receptor-mediated PLC activation. Neomycin, a commonly used phospholipase C blocker, failed to block the receptor-induced PLCd1PH-GFP translocation, indicating that neomycin is unable to block PLC-mediated PtdIns(4,5)P2 hydrolysis. However, in the absence of PLCd1PH-GFP expression, neomycin abolished the receptor-induced hydrolysis of PtdIns(4,5)P2 by PLC.

CONCLUSION

Although PLCd1PH and neomycin bind to PtdIns(4,5)P2 in a similar way, they have distinct effects on receptor-mediated activation of PLC and PtdIns(4,5)P2 hydrolysis.

Authors+Show Affiliations

Department of Pharmacology, Hebei Medical University, Shijiazhuang 050017, China.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

16297348

Citation

Wang, Chuan, et al. "Binding of PLCdelta1PH-GFP to PtdIns(4,5)P2 Prevents Inhibition of Phospholipase C-mediated Hydrolysis of PtdIns(4,5)P2 By Neomycin." Acta Pharmacologica Sinica, vol. 26, no. 12, 2005, pp. 1485-91.
Wang C, Du XN, Jia QZ, et al. Binding of PLCdelta1PH-GFP to PtdIns(4,5)P2 prevents inhibition of phospholipase C-mediated hydrolysis of PtdIns(4,5)P2 by neomycin. Acta Pharmacol Sin. 2005;26(12):1485-91.
Wang, C., Du, X. N., Jia, Q. Z., & Zhang, H. L. (2005). Binding of PLCdelta1PH-GFP to PtdIns(4,5)P2 prevents inhibition of phospholipase C-mediated hydrolysis of PtdIns(4,5)P2 by neomycin. Acta Pharmacologica Sinica, 26(12), 1485-91.
Wang C, et al. Binding of PLCdelta1PH-GFP to PtdIns(4,5)P2 Prevents Inhibition of Phospholipase C-mediated Hydrolysis of PtdIns(4,5)P2 By Neomycin. Acta Pharmacol Sin. 2005;26(12):1485-91. PubMed PMID: 16297348.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Binding of PLCdelta1PH-GFP to PtdIns(4,5)P2 prevents inhibition of phospholipase C-mediated hydrolysis of PtdIns(4,5)P2 by neomycin. AU - Wang,Chuan, AU - Du,Xiao-Na, AU - Jia,Qing-Zhong, AU - Zhang,Hai-Lin, PY - 2005/11/22/pubmed PY - 2007/9/5/medline PY - 2005/11/22/entrez SP - 1485 EP - 91 JF - Acta pharmacologica Sinica JO - Acta Pharmacol. Sin. VL - 26 IS - 12 N2 - AIM: To investigate the effects of the pleckstrin homology (PH) domain of phospholipase C(delta1) (PLC(delta1)PH) on inhibition of phospholipase C (PLC)-mediated hydrolysis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] by neomycin. METHODS: A fusion construct of green fluorescent protein (GFP) and PLC (delta1)PH (PLC(delta1)PH-GFP), which is known to bind PtdIns(4,5)P2 specifically, together with laser-scanning confocal microscopy, was used to trace PtdIns(4,5)P2 translocation. RESULTS: Stimulation of the type 1 muscarinic receptor and the bradykinin 2 receptor induced a reversible PLC(delta1)PH-GFP translocation from the membrane to the cytosol in COS-7 cells. PLC inhibitor U73122 blocked the translocation. Wortmannin, a known PtdIns kinase inhibitor, did not affect the translocation induced by ACh, but blocked recovery after translocation, indicating that PtdIns(4,5)P2 hydrolysis occurs through receptor-mediated PLC activation. Neomycin, a commonly used phospholipase C blocker, failed to block the receptor-induced PLCd1PH-GFP translocation, indicating that neomycin is unable to block PLC-mediated PtdIns(4,5)P2 hydrolysis. However, in the absence of PLCd1PH-GFP expression, neomycin abolished the receptor-induced hydrolysis of PtdIns(4,5)P2 by PLC. CONCLUSION: Although PLCd1PH and neomycin bind to PtdIns(4,5)P2 in a similar way, they have distinct effects on receptor-mediated activation of PLC and PtdIns(4,5)P2 hydrolysis. SN - 1671-4083 UR - https://www.unboundmedicine.com/medline/citation/16297348/Binding_of_PLCdelta1PH_GFP_to_PtdIns_45_P2_prevents_inhibition_of_phospholipase_C_mediated_hydrolysis_of_PtdIns_45_P2_by_neomycin_ L2 - http://dx.doi.org/10.1111/j.1745-7254.2005.00223.x DB - PRIME DP - Unbound Medicine ER -