Effects of calcium on recombinant bovine chromogranin A

R. H. Angeletti, G. Ali, N. Shen, P. Gee, Edward Nieves

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Bovine chromogranin A, the acidic calcium-binding protein characteristic of endocrine secretory vesicles, has been expressed in Escherichia coli using the pET3a vector system under T7 polymerase control. The expressed protein is located in the bacterial cytosol and can be purified from bacterial proteins by a heat treatment step, followed by gel filtration, anion-exchange, and reversed-phase chromatography. The purified recombinant chromogranin A has an apparent M(r) of ca. 72,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in spite of its 432-amino acid polypeptide chain, consistent with observations on natural chromogranin A. The primary structure has been confirmed by mass spectral analysis of tryptic peptides, by Edman degradation of the intact protein, and by immunoreactivity with sequence-specific antibodies. Analysis by circular dichroism spectroscopy shows pH- and concentration-dependent spectra. The spectra are Ca2+-dependent from 5 to 40 μM.

Original languageEnglish (US)
Pages (from-to)1604-1612
Number of pages9
JournalProtein Science
Volume1
Issue number12
StatePublished - 1992

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Chromogranin A
Calcium
Circular dichroism spectroscopy
Peptides
Calcium-Binding Proteins
Bacterial Proteins
Secretory Vesicles
Reverse-Phase Chromatography
Circular Dichroism
Chromatography
Electrophoresis
Sodium Dodecyl Sulfate
Cytosol
Spectrum analysis
Escherichia coli
Proteolysis
Gel Chromatography
Anions
Polyacrylamide Gel Electrophoresis
Spectrum Analysis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Angeletti, R. H., Ali, G., Shen, N., Gee, P., & Nieves, E. (1992). Effects of calcium on recombinant bovine chromogranin A. Protein Science, 1(12), 1604-1612.

Effects of calcium on recombinant bovine chromogranin A. / Angeletti, R. H.; Ali, G.; Shen, N.; Gee, P.; Nieves, Edward.

In: Protein Science, Vol. 1, No. 12, 1992, p. 1604-1612.

Research output: Contribution to journalArticle

Angeletti, RH, Ali, G, Shen, N, Gee, P & Nieves, E 1992, 'Effects of calcium on recombinant bovine chromogranin A', Protein Science, vol. 1, no. 12, pp. 1604-1612.
Angeletti RH, Ali G, Shen N, Gee P, Nieves E. Effects of calcium on recombinant bovine chromogranin A. Protein Science. 1992;1(12):1604-1612.
Angeletti, R. H. ; Ali, G. ; Shen, N. ; Gee, P. ; Nieves, Edward. / Effects of calcium on recombinant bovine chromogranin A. In: Protein Science. 1992 ; Vol. 1, No. 12. pp. 1604-1612.
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