Abstract
Sortilin acts as a cell surface receptor for pro-neurotrophins (pro-NT) that upon complex formation with the p75 neurotrophin receptor (p75(NTR)) is able to signal neuronal cell death. Here we screened a sortilin peptide library comprising 16-mer overlapping sequences for binding of the pro-domains of nerve growth factor and brain-derived neurotrophic factor. We find that a linear surface-exposed sequence, (163)RIFRSSDFAKNF(174), constitutes an important pro-NT binding epitope in sortilin. Systematic mutational analysis revealed residues Arg(163), Phe(165), Arg(166), and Phe(170) to be critical for the interaction. Expression of a sortilin mutant in which these four amino acids were substituted by alanines disrupted pro-NT binding without affecting receptor heterodimerization with p75(NTR) or binding of ligands that selectively engages the centrally located tunnel in the beta-propeller of sortilin. We furthermore demonstrate that a peptide comprising the ligand-binding epitope can prevent pro-NT-induced apoptosis in RN22 schwannoma cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Vesicular Transport / chemistry*
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Adaptor Proteins, Vesicular Transport / genetics
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Adaptor Proteins, Vesicular Transport / immunology
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Adaptor Proteins, Vesicular Transport / metabolism*
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Amino Acid Sequence
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Amino Acid Substitution
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Apoptosis
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Base Sequence
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Binding Sites / genetics
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Cell Line
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DNA Primers / genetics
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Dimerization
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Epitopes / chemistry
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Epitopes / genetics
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Epitopes / metabolism
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Humans
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In Vitro Techniques
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Ligands
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Models, Molecular
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Nerve Growth Factors / metabolism*
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Nerve Tissue Proteins / chemistry
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Nerve Tissue Proteins / metabolism
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Protein Binding
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Protein Precursors / metabolism
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Receptors, Nerve Growth Factor / chemistry
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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 / immunology
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Recombinant Proteins / metabolism
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Transfection
Substances
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Adaptor Proteins, Vesicular Transport
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DNA Primers
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Epitopes
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Ligands
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NGFR protein, human
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Nerve Growth Factors
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Nerve Tissue Proteins
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Protein Precursors
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Receptors, Nerve Growth Factor
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Recombinant Proteins
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sortilin