Characterization of metastable intermediates of enzymatic peroxidation of NAD+ by on-line capillary electrophoresis/electrospray ionization mass spectrometry

• Published in: Journal of mass spectrometry. Vol. 31; no. 2; pp. 193 - 198
• Main Authors: Zhao, Zhongxi (Zack), Udseth, Harold R., Smith, Richard D.
• Format: Journal Article
• Language: English
• Published: Sussex John Wiley & Sons, Ltd 01.02.1996; Wiley
• Subjects: alcohol dehydrogenase; Analytical, structural and metabolic biochemistry; Biological and medical sciences; capillary electrophoresis; capillary isotachophoresis; electrospray ionization; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; nicotinamide adenine dinucleotide (NAD+); Oxidoreductases; Capillary column; Reaction intermediate; On line; Dinucleotide; Alcohol dehydrogenase; Electrospray; Horse; Reaction mechanism; Electrophoresis; Ungulata; Peroxidation; Collisional activation; Enzyme; Coupled method; Spraying; Isotachophoresis; Hydrogen Peroxides; NAD; Vertebrata; Mammalia; Electric field; Mass spectrometry MS/MS; Oxidoreductases; Perissodactyla; Enzymatic reaction
• ISSN: 1076-5174; 1096-9888
• DOI: 10.1002/(SICI)1096-9888(199602)31:2<193::AID-JMS282>3.0.CO;2-7

The unstable intermediate products formed from the degradation of nicotinamide adenine dinucleotide (NAD+) catalyzed by the reaction of equine liver alcohol dehydrogenase with hydrogen peroxide were studied. Reversed anionic capillary electrophoresis (CE)/electrospray ionization mass spectrometry (ESI‐MS) was used to separate the reaction mixture and to monitor the reaction process, and to provide both molecular weight and structural information for the interaction intermediates. The mass spectra of the reaction intermediates provide support for the reaction mechanism previously proposed by Favilla et al. The present results also indicate that a different reaction intermediate may also be involved in the interaction, and contributes to the reaction. The results illustrate the utility of combined CE and ESI‐MS for the on‐line study of biochemical processes.