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A human single-chain Fv intrabody blocks aberrant cellular effects of overexpressed α-synuclein
by
Sierks, Michael R.
, Zhou, Chun
, Messer, Anne
, Emadi, Sharareh
in
alpha-Synuclein
/ Animals
/ Antibody Specificity
/ Cell Adhesion - drug effects
/ Cell Line
/ Cercopithecus aethiops
/ Detergents - pharmacology
/ Gene Expression
/ Humans
/ Immunoglobulin Fragments - immunology
/ Immunoglobulin Fragments - pharmacology
/ Nerve Tissue Proteins - antagonists & inhibitors
/ Nerve Tissue Proteins - genetics
/ Nerve Tissue Proteins - immunology
/ Nerve Tissue Proteins - metabolism
/ Protein Denaturation - drug effects
/ Solubility - drug effects
/ Synucleins
2004
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A human single-chain Fv intrabody blocks aberrant cellular effects of overexpressed α-synuclein
by
Sierks, Michael R.
, Zhou, Chun
, Messer, Anne
, Emadi, Sharareh
in
alpha-Synuclein
/ Animals
/ Antibody Specificity
/ Cell Adhesion - drug effects
/ Cell Line
/ Cercopithecus aethiops
/ Detergents - pharmacology
/ Gene Expression
/ Humans
/ Immunoglobulin Fragments - immunology
/ Immunoglobulin Fragments - pharmacology
/ Nerve Tissue Proteins - antagonists & inhibitors
/ Nerve Tissue Proteins - genetics
/ Nerve Tissue Proteins - immunology
/ Nerve Tissue Proteins - metabolism
/ Protein Denaturation - drug effects
/ Solubility - drug effects
/ Synucleins
2004
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A human single-chain Fv intrabody blocks aberrant cellular effects of overexpressed α-synuclein
by
Sierks, Michael R.
, Zhou, Chun
, Messer, Anne
, Emadi, Sharareh
in
alpha-Synuclein
/ Animals
/ Antibody Specificity
/ Cell Adhesion - drug effects
/ Cell Line
/ Cercopithecus aethiops
/ Detergents - pharmacology
/ Gene Expression
/ Humans
/ Immunoglobulin Fragments - immunology
/ Immunoglobulin Fragments - pharmacology
/ Nerve Tissue Proteins - antagonists & inhibitors
/ Nerve Tissue Proteins - genetics
/ Nerve Tissue Proteins - immunology
/ Nerve Tissue Proteins - metabolism
/ Protein Denaturation - drug effects
/ Solubility - drug effects
/ Synucleins
2004
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A human single-chain Fv intrabody blocks aberrant cellular effects of overexpressed α-synuclein
Journal Article
A human single-chain Fv intrabody blocks aberrant cellular effects of overexpressed α-synuclein
2004
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Overview
alpha-Synuclein (alpha-syn) has been identified as the major component of Lewy bodies that characterize neurodegenerative synucleinopathies, including Parkinson's disease. Overexpression of alpha-syn, and prefibrillar alpha-syn oligomers, has been implicated in these pathologies; therefore, prevention of prefibril accumulation, and inhibition of other aberrant effects of overexpressed alpha-syn, could provide novel treatments. Here, we have selected a human single-chan Fv (scFv) antibody, D10, that binds human monomeric wild-type alpha-syn. We demonstrate, by retargeting assays and coimmunoprecipitation, that the D10 scFv is a specific and efficient intracellular antibody (intrabody). By transfecting the D10 scFv gene into an HEK 293 cell line that overexpresses wild-type alpha-syn, we show that the D10 intrabody stabilizes detergent-soluble monomeric alpha-syn and inhibits the formation of detergent-insoluble high-molecular-weight alpha-syn species. In addition, the D10 intrabody ameliorates the decreased cell adhesion that characterizes the alpha-syn-overexpressing cells. Given the important role of alpha-syn pathology, and the facility with which intrabodies can be further engineered in vitro, anti-alpha-syn intrabodies may represent novel molecular therapeutics for synucleinopathies, with implications for other neurodegenerative disorders caused by misfolded accumulated proteins.
Subject
/ Animals
/ Cell Adhesion - drug effects
/ Humans
/ Immunoglobulin Fragments - immunology
/ Immunoglobulin Fragments - pharmacology
/ Nerve Tissue Proteins - antagonists & inhibitors
/ Nerve Tissue Proteins - genetics
/ Nerve Tissue Proteins - immunology
/ Nerve Tissue Proteins - metabolism
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