Abstract
Lynx1 expresses in the central nervous system and plays important role in a regulation of nicotinic acetylcholine receptors. Successful milligram-quantitive expression of ws-Lynx1 was achieved only in the case of its production in the form of cytoplasm inclusion bodies. Different conditions of ws-Lynx1 refolding for yield optimization were performed. The obtained recombinant protein was characterized by means of mass spectrometry and CD spectroscopy. The binding experiments on the nAChRs from Torpedo californica membranes revealed that ws-Lynxl is biologically active and blocks muscle nAChR with IC50-20-30 microM.
MeSH terms
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Adaptor Proteins, Signal Transducing
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Amino Acid Sequence
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Animals
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Base Sequence
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Bungarotoxins / antagonists & inhibitors
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Electric Organ / chemistry
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Electric Organ / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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GPI-Linked Proteins / biosynthesis*
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GPI-Linked Proteins / chemistry
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GPI-Linked Proteins / pharmacology
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Gene Expression
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Genetic Vectors
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Humans
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Molecular Sequence Data
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Neurotransmitter Agents / biosynthesis*
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Neurotransmitter Agents / chemistry
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Neurotransmitter Agents / pharmacology
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Protein Structure, Tertiary
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Receptors, Nicotinic / metabolism*
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Recombinant Proteins / biosynthesis*
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Recombinant Proteins / chemistry
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Recombinant Proteins / pharmacology
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Solubility
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Torpedo
Substances
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Adaptor Proteins, Signal Transducing
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Bungarotoxins
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GPI-Linked Proteins
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LYNX1 protein, human
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Neurotransmitter Agents
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Receptors, Nicotinic
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Recombinant Proteins