Abstract
Nerve growth factor (NGF) is involved in the development and maintenance of the nervous system. NGF binds with high affinity to the extracellular region of the tyrosine kinase receptor TrkA. This domain comprises leucine and cysteine rich motifs, followed by two immunoglobulin like (Ig-like) domains. We describe the expression and purification of recombinant Ig-like domains. Fluorescence and circular dichroism spectroscopy show that the protein is folded into a compact globular structure and contains mainly beta-sheet secondary structure. Recombinant protein binds to NGF and can inhibit NGF bioactivity both in vitro and in vivo.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Binding Sites / genetics
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Binding, Competitive
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Biotechnology
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Circular Dichroism
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Humans
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Immunoglobulins / chemistry
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Immunoglobulins / genetics*
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Immunoglobulins / metabolism*
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In Vitro Techniques
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Male
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Molecular Sequence Data
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Nerve Growth Factors / antagonists & inhibitors
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Nerve Growth Factors / metabolism*
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PC12 Cells
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Protein Folding
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Protein Structure, Secondary
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Proto-Oncogene Proteins / chemistry
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Proto-Oncogene Proteins / genetics*
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Proto-Oncogene Proteins / metabolism*
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Rats
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Rats, Wistar
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Receptor Protein-Tyrosine Kinases / chemistry
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Receptor Protein-Tyrosine Kinases / genetics*
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Receptor Protein-Tyrosine Kinases / metabolism*
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Receptor, trkA
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Receptors, Nerve Growth Factor / chemistry
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Receptors, Nerve Growth Factor / genetics*
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Receptors, Nerve Growth Factor / metabolism*
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Spectrometry, Fluorescence
Substances
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Immunoglobulins
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Nerve Growth Factors
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Proto-Oncogene Proteins
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Receptors, Nerve Growth Factor
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Recombinant Proteins
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Receptor Protein-Tyrosine Kinases
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Receptor, trkA