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: Research - peer-reviewArticle

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

    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

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    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",
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    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

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    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

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