Isolation and identification of intact chromogranin A and two N-terminal processing products, vasostatin I and II, from bovine adrenal medulla chromaffin granules by chromatographic and mass spectrometric methods

Chromogranin A (CGA) is the most abundant protein of the bovine adrenal medulla and plays an important role as precursor protein of several peptides that act as modulators for endocrine cell secretory activity. Furthermore, it is presumed to play a role in the targeting of peptide hormones and neuro...

Full description

Saved in:
Bibliographic Details
Published in:Neuropeptides (Edinburgh) 1997-06, Vol.31 (3), p.273-280
Main Authors: Bauer, S.H.J, Zhang, X.Y, Liang, F, De Potter, W.P, Claeys, M, Przybylski, M
Format: Article
Language:English
Subjects:
Adrenals. Interrenals
Adrenomedullary hormones. Regulation
Amino Acid Sequence
Animals
Biological and medical sciences
Cattle
Chromaffin Cells - chemistry
Chromaffin Cells - ultrastructure
Chromaffin Granules - chemistry
Chromaffin Granules - metabolism
Chromatography, High Pressure Liquid
Chromogranin A
Chromogranins - analysis
Chromogranins - genetics
Chromogranins - metabolism
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Mass Spectrometry
Molecular Sequence Data
Peptide Fragments - analysis
Peptide Fragments - genetics
Vasodilator Agents - analysis
Vasodilator Agents - metabolism
Vertebrates: endocrinology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Chromogranin A (CGA) is the most abundant protein of the bovine adrenal medulla and plays an important role as precursor protein of several peptides that act as modulators for endocrine cell secretory activity. Furthermore, it is presumed to play a role in the targeting of peptide hormones and neurotransmitters to granules of the regulated pathway. However, its complete primary structure and proteolytic processing have not yet been identified. This study describes a rapid and efficient procedure for the high yield isolation of bovine CGA and its N-terminal processing products, vasostatin I and II. Using the lysate from bovine adrenal medulla chromaffin granules, the soluble proteins were purified by three consecutive HPLC steps, thereby avoiding the use of buffer solutions. The protein fractions were isolated and characterized by SDS-PAGE and Western blot analysis as well as by mass spectrometry. In the latter analysis, the efficiency of matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) was demonstrated, enabling the unequivocal and sensitive characterization of proteins from crude mixtures. Sufficient amounts of pure protein were obtained by the present procedure to form the basis for detailed structural studies by spectroscopic methods and X-ray crystallography.
ISSN:0143-4179
1532-2785
DOI:10.1016/S0143-4179(97)90059-7