title: Immunoglobulin-like domains define the nerve growth factor binding site of the TrkA receptor.
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.
authors.affiliation: Department of Medicine (Care of the Elderly), Bristol Royal Infirmary, UK. p.h.holden@bris.ac.uk
authors.forename: P H
authors.initials: PH
authors.lastname: Holden
authors.forename: V
authors.initials: V
authors.lastname: Asopa
authors.forename: A G
authors.initials: AG
authors.lastname: Robertson
authors.forename: A R
authors.initials: AR
authors.lastname: Clarke
authors.forename: S
authors.initials: S
authors.lastname: Tyler
authors.forename: G S
authors.initials: GS
authors.lastname: Bennett
authors.forename: S D
authors.initials: SD
authors.lastname: Brain
authors.forename: G K
authors.initials: GK
authors.lastname: Wilcock
authors.forename: S J
authors.initials: SJ
authors.lastname: Allen
authors.forename: S K
authors.initials: SK
authors.lastname: Smith
authors.forename: D
authors.initials: D
authors.lastname: Dawbarn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D001667:Binding, Competitive; D001709:Biotechnology; D002942:Circular Dichroism; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016716:PC12 Cells; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020917:Receptor, trkA; D017475:Receptors, Nerve Growth Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D007136:Immunoglobulins; D009414:Nerve Growth Factors; D011518:Proto-Oncogene Proteins; D017475:Receptors, Nerve Growth Factor; D011994:Recombinant Proteins; D020794:Receptor Protein-Tyrosine Kinases; D020917:Receptor, trkA