Extracellular domain of neurotrophin receptor trkB: disulfide structure, N-glycosylation sites, and ligand binding

Arch Biochem Biophys. 1995 Sep 10;322(1):256-64. doi: 10.1006/abbi.1995.1460.

Abstract

An extracellular domain of a human neurotrophin receptor trkB was expressed in Chinese hamster ovary cells and isolated as a glycoprotein possessing binding activity for brain-derived neurotrophic factor. The extracellular domain contains 398 amino acids and has a molecular weight of 60.6 kDa according to laser desorption mass spectrometry, indicating that the extracellular domain of trkB contains 33.3% carbohydrate moieties. Six disulfide linkages were determined to be Cys1-Cys7, Cys5-Cys14, Cys121-Cys145, Cys123-Cys163, Cys187-Cys235, and Cys271-Cys314, respectively. Cys300 was detected as a free sulfhydryl residue. Cysteine clusters 1 and 2 located in the N-terminal domain possess a similar type of disulfide structure and two other disulfide bonds in the C-terminal region are homologous to that of the Ig-like C2 domain. Among 12 potential N-linked glycosylation sites proposed in the soluble domain of trkB, 10 sites are actually glycosylated.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • CHO Cells
  • Cricetinae
  • Cross-Linking Reagents
  • Disulfides / chemistry
  • Glycosylation
  • Humans
  • Ligands
  • Molecular Sequence Data
  • Molecular Structure
  • Molecular Weight
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Mapping
  • Receptor, Ciliary Neurotrophic Factor
  • Receptors, Nerve Growth Factor / chemistry*
  • Receptors, Nerve Growth Factor / genetics
  • Receptors, Nerve Growth Factor / metabolism
  • Solubility

Substances

  • Cross-Linking Reagents
  • Disulfides
  • Ligands
  • Peptide Fragments
  • Receptor, Ciliary Neurotrophic Factor
  • Receptors, Nerve Growth Factor