Document Detail


Evidence that the chromogranin B fragment 368-417 extracted from a pheochromocytoma is phosphorylated.
MedLine Citation:
PMID:  11514034     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A rabbit antiserum was raised against a synthetic peptide corresponding to residues 403 to 417 of human chromogranin B. This peptide was chosen to match the potential C-terminal end of a putative proteolytic fragment of the protein located between dibasic doublets in positions 366-367 and in positions 418-419 of the precursor. A radioimmunoassay based on this antiserum was developed and used to detect the protein or a fragment thereof in a pheochromocytoma tumor extract. One fragment was purified to homogeneity by successive reverse-phase HPLC chromatographies. The N-terminal sequence established by automated Edman degradation, was N-Y-P-S-L-E-L-D-K-M-A-H-G-Y-G-E-E-S-E-E-E-R corresponding to the 368-389 sequence of human chromogranin B. Taking into account the specificity of the antiserum used for peptide identification and alignment with the precursor sequence, we deduced that the purified peptide was chromogranin B (368-417) and represented a new peptide generated by limited proteolysis of chromogranin B. Combining electrospray mass-spectrometry and enzymatic dephosphorylation, we demonstrated that this peptide was phosphorylated.
Authors:
H Dahma; P Gourlet; A Vandermeers; M C Vandermeers-Piret; P Robberecht
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Peptides     Volume:  22     ISSN:  0196-9781     ISO Abbreviation:  Peptides     Publication Date:  2001 Sep 
Date Detail:
Created Date:  2001-08-21     Completed Date:  2001-12-21     Revised Date:  2007-04-17    
Medline Journal Info:
Nlm Unique ID:  8008690     Medline TA:  Peptides     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1491-9     Citation Subset:  IM    
Affiliation:
Department of Biochemistry and Nutrition, Medical School, Université Libre de Bruxelles, B-1070, Brussels, Belgium.
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MeSH Terms
Descriptor/Qualifier:
Adrenal Gland Neoplasms / chemistry*
Adult
Amino Acid Sequence
Animals
Chromatography, High Pressure Liquid
Chromogranin B
Chromogranins / analysis*,  biosynthesis,  chemistry,  genetics,  metabolism
Humans
Male
Neoplasm Proteins / isolation & purification*,  metabolism
Peptide Fragments / analysis*,  biosynthesis,  chemical synthesis,  chemistry,  genetics
Pheochromocytoma / chemistry*
Phosphorylation
Rabbits / immunology
Radioimmunoassay
Sensitivity and Specificity
Sequence Alignment
Sequence Homology, Amino Acid
Spectrometry, Mass, Electrospray Ionization
Chemical
Reg. No./Substance:
0/CHGB protein, human; 0/Chromogranin B; 0/Chromogranins; 0/Neoplasm Proteins; 0/Peptide Fragments; 0/chromogranin B (368-417)

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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