Chromogranin a epitopes: Clues from synthetic peptides and peptide mapping

B. M. Gill, J. A. Barbosa, R. Hogue-Angeletti, N. Varki, D. T. O'Connor

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Chromogranin A (CgA) is a 48 kDa acidic protein in neuroendocrine secretory vesicles whose primary structure is now known. We used synthetic peptides, synthetic peptide antisera, intact molecule antisera, chymotryptic peptide mapping, microsequencing, immunoblotting, and immunoprecipitation to probe the location of immunodominant domains within the CgA molecule. Polyclonal anti mid-molecule, anti N-terminal and anti C-terminal antibodies specifically visualized CgA (both bovine and human) in one and two dimensional immunoblots of adrenal chromaffin vesicles, and the stain CgA fragments further suggested bidirectional (both N- and C-terminal) cleavage or processing of CgA. Anti intact CgA immunoblotting of HPLC-separated peptides from chymotrypsin-digested bovine CgA revealed several strongly immunoreactive internal peptides, two of which were positioned by N-terminal amino acid sequencing: CgA91ff. and CgA197ff.. A single synthetic peptide (CgA79-113) was recognized by three antibodies developed against the intact CgA molecule: two polyclonal rabbit antisera as well as a monoclonal mouse antibody. Not all antigenicity algorithm-predicted domains were immunogenic, suggesting that some of these predicted domains may not be accessible. Polyclonal anti mid-molecule, anti N- and anti C-terminal synthetic peptide antisera specifically immunoprecipitated 125I-labeled bovine CgA from aqueous solution; mid-molecule antisera precipitated substantially greater amounts than terminal antisera. The immunoprecipitation results suggested exposed terminal as well as interior hydrophilic epitopes in the molecule in its intact, native conformation. 125I-human CgA was best precipitated by anti N-terminal antisera, consistent with greatest interspecies sequence conservation at the N-terminus of CgA. The terminal antisera reacted immunohistochemically in a granular pattern with adrenal medullary chromaffin cells (but not adrenal cortical cells) and pancreatic islet cells (but not pancreatic exocrine acini). Thus, synthetic and chymotryptic peptides yielded novel and specific insights into the structure, conformation, vesicular processing and interior immunodominant domains of CgA.

Original languageEnglish (US)
Pages (from-to)105-118
Number of pages14
JournalNeuropeptides
Volume21
Issue number2
DOIs
StatePublished - 1992

Fingerprint

Chromogranins
Chromogranin A
Peptide Mapping
Epitopes
Immune Sera
Peptides
Molecules
Immunodominant Epitopes
Islets of Langerhans
Immunoprecipitation
Immunoblotting
Conformations
Antibodies
Chromaffin Cells
Protein Sequence Analysis
Secretory Vesicles
Chymotrypsin
Processing
Conservation
Coloring Agents

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Endocrinology, Diabetes and Metabolism
  • Clinical Neurology
  • Neuroscience(all)
  • Cellular and Molecular Neuroscience

Cite this

Gill, B. M., Barbosa, J. A., Hogue-Angeletti, R., Varki, N., & O'Connor, D. T. (1992). Chromogranin a epitopes: Clues from synthetic peptides and peptide mapping. Neuropeptides, 21(2), 105-118. https://doi.org/10.1016/0143-4179(92)90521-W

Chromogranin a epitopes : Clues from synthetic peptides and peptide mapping. / Gill, B. M.; Barbosa, J. A.; Hogue-Angeletti, R.; Varki, N.; O'Connor, D. T.

In: Neuropeptides, Vol. 21, No. 2, 1992, p. 105-118.

Research output: Contribution to journalArticle

Gill, BM, Barbosa, JA, Hogue-Angeletti, R, Varki, N & O'Connor, DT 1992, 'Chromogranin a epitopes: Clues from synthetic peptides and peptide mapping', Neuropeptides, vol. 21, no. 2, pp. 105-118. https://doi.org/10.1016/0143-4179(92)90521-W
Gill, B. M. ; Barbosa, J. A. ; Hogue-Angeletti, R. ; Varki, N. ; O'Connor, D. T. / Chromogranin a epitopes : Clues from synthetic peptides and peptide mapping. In: Neuropeptides. 1992 ; Vol. 21, No. 2. pp. 105-118.
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