Influence of lysophospholipid hydrolysis by the catalytic domain of neuropathy target esterase on the fluidity of bilayer lipid membranes

Aaron J. Greiner, Rudy J. Richardson, R. Mark Worden, Robert Y. Ofoli

Research output: Contribution to journalArticle

  • 10 Citations

Abstract

Neuropathy target esterase (NTE) is an integral membrane protein localized in the endoplasmic reticulum in neurons. Irreversible inhibition of NTE by certain organophosphorus compounds produces a paralysis known as organophosphorus compound-induced delayed neuropathy. In vitro, NTE has phospholipase/lysophospholipase activity that hydrolyses exogenously added single-chain lysophospholipids in preference to dual-chain phospholipids, and NTE mutations have been associated with motor neuron disease. NTE's physiological role is not well understood, although recent studies suggest that it may control the cytotoxic accumulation of lysophospholipids in membranes. We used the NTE catalytic domain (NEST) to hydrolyze palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (p-lysoPC) to palmitic acid in bilayer membranes comprising 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and the fluorophore 1-oleoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (NBD-PC). Translational diffusion coefficients (DL) in supported bilayer membranes were measured by fluorescence recovery after pattern photobleaching (FRAPP). The average DL for DOPC/p-lysoPC membranes without NEST was 2.44μm2s-1±0.09; the DL for DOPC/p-lysoPC membranes containing NEST and diisopropylphosphorofluoridate, an inhibitor, was nearly identical at 2.45±0.08. By contrast, the DL for membranes comprising NEST, DOPC, and p-lysoPC was 2.28±0.07, significantly different from the system with inhibited NEST, due to NEST hydrolysis. Likewise, a system without NEST containing the amount of palmitic acid that would have been produced by NEST hydrolysis of p-lysoPC was identical at 2.26±0.06. These results indicate that NTE's catalytic activity can alter membrane fluidity.

LanguageEnglish (US)
Pages1533-1539
Number of pages7
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1798
Issue number8
DOIs
StatePublished - Aug 2010

Profile

Lysophospholipids
Membrane Fluidity
Lipid bilayers
Fluidity
Lipid Bilayers
Membrane Lipids
Phosphorylcholine
Hydrolysis
Catalytic Domain
Membranes
Organophosphorus Compounds
Palmitic Acid
Neurons
Lysophospholipase
Fluorescence Recovery After Photobleaching
Isoflurophate
Motor Neuron Disease
Phospholipases
Photobleaching
Fluorophores

Keywords

  • Bilayer membrane fluidity
  • Lysophospholipid hydrolysis
  • Neuropathy target esterase (NTE)
  • Neuropathy target esterase catalytic domain (NEST)
  • Organophosphorus inhibition
  • Supported bilayer lipid membrane

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Biophysics

Cite this

@article{14b0eee570fb42ee87db9ac83bcd6881,
title = "Influence of lysophospholipid hydrolysis by the catalytic domain of neuropathy target esterase on the fluidity of bilayer lipid membranes",
abstract = "Neuropathy target esterase (NTE) is an integral membrane protein localized in the endoplasmic reticulum in neurons. Irreversible inhibition of NTE by certain organophosphorus compounds produces a paralysis known as organophosphorus compound-induced delayed neuropathy. In vitro, NTE has phospholipase/lysophospholipase activity that hydrolyses exogenously added single-chain lysophospholipids in preference to dual-chain phospholipids, and NTE mutations have been associated with motor neuron disease. NTE's physiological role is not well understood, although recent studies suggest that it may control the cytotoxic accumulation of lysophospholipids in membranes. We used the NTE catalytic domain (NEST) to hydrolyze palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (p-lysoPC) to palmitic acid in bilayer membranes comprising 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and the fluorophore 1-oleoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (NBD-PC). Translational diffusion coefficients (DL) in supported bilayer membranes were measured by fluorescence recovery after pattern photobleaching (FRAPP). The average DL for DOPC/p-lysoPC membranes without NEST was 2.44μm2s-1±0.09; the DL for DOPC/p-lysoPC membranes containing NEST and diisopropylphosphorofluoridate, an inhibitor, was nearly identical at 2.45±0.08. By contrast, the DL for membranes comprising NEST, DOPC, and p-lysoPC was 2.28±0.07, significantly different from the system with inhibited NEST, due to NEST hydrolysis. Likewise, a system without NEST containing the amount of palmitic acid that would have been produced by NEST hydrolysis of p-lysoPC was identical at 2.26±0.06. These results indicate that NTE's catalytic activity can alter membrane fluidity.",
keywords = "Bilayer membrane fluidity, Lysophospholipid hydrolysis, Neuropathy target esterase (NTE), Neuropathy target esterase catalytic domain (NEST), Organophosphorus inhibition, Supported bilayer lipid membrane",
author = "Greiner, {Aaron J.} and Richardson, {Rudy J.} and Worden, {R. Mark} and Ofoli, {Robert Y.}",
year = "2010",
month = "8",
doi = "10.1016/j.bbamem.2010.03.015",
language = "English (US)",
volume = "1798",
pages = "1533--1539",
journal = "Biochimica et Biophysica Acta - Biomembranes",
issn = "0005-2736",
publisher = "Elsevier",
number = "8",

}

TY - JOUR

T1 - Influence of lysophospholipid hydrolysis by the catalytic domain of neuropathy target esterase on the fluidity of bilayer lipid membranes

AU - Greiner,Aaron J.

AU - Richardson,Rudy J.

AU - Worden,R. Mark

AU - Ofoli,Robert Y.

PY - 2010/8

Y1 - 2010/8

N2 - Neuropathy target esterase (NTE) is an integral membrane protein localized in the endoplasmic reticulum in neurons. Irreversible inhibition of NTE by certain organophosphorus compounds produces a paralysis known as organophosphorus compound-induced delayed neuropathy. In vitro, NTE has phospholipase/lysophospholipase activity that hydrolyses exogenously added single-chain lysophospholipids in preference to dual-chain phospholipids, and NTE mutations have been associated with motor neuron disease. NTE's physiological role is not well understood, although recent studies suggest that it may control the cytotoxic accumulation of lysophospholipids in membranes. We used the NTE catalytic domain (NEST) to hydrolyze palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (p-lysoPC) to palmitic acid in bilayer membranes comprising 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and the fluorophore 1-oleoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (NBD-PC). Translational diffusion coefficients (DL) in supported bilayer membranes were measured by fluorescence recovery after pattern photobleaching (FRAPP). The average DL for DOPC/p-lysoPC membranes without NEST was 2.44μm2s-1±0.09; the DL for DOPC/p-lysoPC membranes containing NEST and diisopropylphosphorofluoridate, an inhibitor, was nearly identical at 2.45±0.08. By contrast, the DL for membranes comprising NEST, DOPC, and p-lysoPC was 2.28±0.07, significantly different from the system with inhibited NEST, due to NEST hydrolysis. Likewise, a system without NEST containing the amount of palmitic acid that would have been produced by NEST hydrolysis of p-lysoPC was identical at 2.26±0.06. These results indicate that NTE's catalytic activity can alter membrane fluidity.

AB - Neuropathy target esterase (NTE) is an integral membrane protein localized in the endoplasmic reticulum in neurons. Irreversible inhibition of NTE by certain organophosphorus compounds produces a paralysis known as organophosphorus compound-induced delayed neuropathy. In vitro, NTE has phospholipase/lysophospholipase activity that hydrolyses exogenously added single-chain lysophospholipids in preference to dual-chain phospholipids, and NTE mutations have been associated with motor neuron disease. NTE's physiological role is not well understood, although recent studies suggest that it may control the cytotoxic accumulation of lysophospholipids in membranes. We used the NTE catalytic domain (NEST) to hydrolyze palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (p-lysoPC) to palmitic acid in bilayer membranes comprising 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and the fluorophore 1-oleoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (NBD-PC). Translational diffusion coefficients (DL) in supported bilayer membranes were measured by fluorescence recovery after pattern photobleaching (FRAPP). The average DL for DOPC/p-lysoPC membranes without NEST was 2.44μm2s-1±0.09; the DL for DOPC/p-lysoPC membranes containing NEST and diisopropylphosphorofluoridate, an inhibitor, was nearly identical at 2.45±0.08. By contrast, the DL for membranes comprising NEST, DOPC, and p-lysoPC was 2.28±0.07, significantly different from the system with inhibited NEST, due to NEST hydrolysis. Likewise, a system without NEST containing the amount of palmitic acid that would have been produced by NEST hydrolysis of p-lysoPC was identical at 2.26±0.06. These results indicate that NTE's catalytic activity can alter membrane fluidity.

KW - Bilayer membrane fluidity

KW - Lysophospholipid hydrolysis

KW - Neuropathy target esterase (NTE)

KW - Neuropathy target esterase catalytic domain (NEST)

KW - Organophosphorus inhibition

KW - Supported bilayer lipid membrane

UR - http://www.scopus.com/inward/record.url?scp=77953727658&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77953727658&partnerID=8YFLogxK

U2 - 10.1016/j.bbamem.2010.03.015

DO - 10.1016/j.bbamem.2010.03.015

M3 - Article

VL - 1798

SP - 1533

EP - 1539

JO - Biochimica et Biophysica Acta - Biomembranes

T2 - Biochimica et Biophysica Acta - Biomembranes

JF - Biochimica et Biophysica Acta - Biomembranes

SN - 0005-2736

IS - 8

ER -