Document Detail

Isolation and identification by gas chromatographic mass spectrometry of the carbonyl-active site of pig kidney diamine oxidase.
MedLine Citation:
PMID:  7978244     Owner:  NLM     Status:  MEDLINE    
An adduct with phenylhydrazine was formed with the purified pig kidney diamine oxidase and in parallel with the l-tyrosine decarboxylase from Streptococcus faecalis. The labeled enzymes were hydrolyzed by chemical hydrolysis and the adducts released by hydrolysis were isolated and identified first in HPLC and successively in GC-MS. Both enzymes gave the same adduct which was identified as the phenylhydrazone of pyridoxal. The isolated adduct had the same retention time as the phenylhydrazone of pyridoxal in HPLC and in gas chromatography and showed the same molecular weight in mass spectrometry when chemical ionization was used and the same fragmentation in mass spectrometry when electronic impact was used. The reported results show that pig kidney diamine oxidase contains pyridoxal in the form of covalently linked pyridoxal phosphate which can be released from the enzyme only by chemical hydrolysis. Pig kidney diamine oxidase is therefore a pyridoxal enzyme such as l-tyrosine decarboxylase as hypothesized in the past but never clearly demonstrated.
F Buffoni
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical biochemistry     Volume:  220     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1994 Jul 
Date Detail:
Created Date:  1994-12-12     Completed Date:  1994-12-12     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  185-91     Citation Subset:  IM    
Department of Pharmacology, University of Florence, Italy.
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MeSH Terms
Amine Oxidase (Copper-Containing) / analysis*
Binding Sites
Chromatography, High Pressure Liquid
Enterococcus faecalis / enzymology
Gas Chromatography-Mass Spectrometry*
Kidney / enzymology*
Phenylhydrazines / chemistry
Pyridoxal Phosphate / analysis
Spectrophotometry, Ultraviolet
Tyrosine Decarboxylase / analysis
Reg. No./Substance:
0/Phenylhydrazines; 100-63-0/phenylhydrazine; 54-47-7/Pyridoxal Phosphate; EC Oxidase (Copper-Containing); EC Decarboxylase
Erratum In:
Anal Biochem 1994 Nov 1;222(2):520

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