Phosphorylation and O-glycosylation sites of human chromogranin A (CGA79-439) from urine of patients with carcinoid tumors

J Biol Chem. 1998 Dec 18;273(51):34087-97. doi: 10.1074/jbc.273.51.34087.

Abstract

Because of their water-soluble properties, chromogranins (CGs) and chromogranin-derived fragments are released together with catecholamines from adrenal chromaffin cells during stress situations and can be detected in the blood by radiochemical and enzyme assays. It is well known that chromogranins can serve as immunocytochemical markers for neuroendocrine tissues and as a diagnostic tool for neuroendocrine tumors. In 1993, large CGA-derived fragments have been shown to be excreted into the urine in patients with carcinoid tumors and the present study deals with the characterization of the post-translational modifications (phosphorylation and O-glycosylation) located along the largest natural CGA-derived fragment CGA79-439. Using mild proteolysis of peptidic material, high performance liquid chromatography, sequencing, and mass spectrometry analysis, six post-translational modifications were detected along the C-terminal CGA-derived fragment CGA79-439. Three O-linked glycosylation sites were located in the core of the protein on Thr163, Thr165, and Thr233, consisting in di-, tri-, and tetrasaccharides. Three phosphorylation sites were located in the middle and C-terminal domain, on serine residues Ser200, Ser252, and Ser315. These modified sites were compared with sequences of others species and discussed in relation with the post-translational modifications that we have reported previously for bovine CGA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biomarkers, Tumor / urine
  • Carcinoid Tumor / secondary
  • Carcinoid Tumor / urine*
  • Cattle
  • Chromatography, High Pressure Liquid
  • Chromogranin A
  • Chromogranins / chemistry*
  • Chromogranins / metabolism*
  • Chromogranins / urine
  • Glycopeptides / chemistry
  • Glycopeptides / isolation & purification
  • Glycosylation
  • Humans
  • Liver Neoplasms / secondary
  • Liver Neoplasms / urine
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Peptide Fragments / ultrastructure
  • Phosphopeptides / chemistry
  • Phosphopeptides / isolation & purification
  • Phosphorylation
  • Protein Processing, Post-Translational*
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Biomarkers, Tumor
  • CHGA protein, human
  • Chromogranin A
  • Chromogranins
  • Glycopeptides
  • Peptide Fragments
  • Phosphopeptides