Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1215674 | Journal of Chromatography B | 2007 | 5 Pages |
Abstract
The reaction from retinal to retinoic acid catalyzed by retinal dehydrogenase on a polyvinylidene difluoride (PVDF) membrane was examined using laser desorption ionization time of flight mass spectrometry (LDI-TOF MS) when the enzyme was separated by non-denaturing two-dimensional electrophoresis (2-DE), transferred onto the membrane, and stained without impairing the enzyme activity. Furthermore, the enzyme was analyzed by de novo sequencing using electrospray ionization tandem mass spectrometry (ESI-MS/MS) after proteins from mouse liver were separated by non-denaturing 2-DE, blotted onto the membrane, and stained. The results indicated that the reported methods could be applied for the direct examination of changes in retinoid catalyzed by enzymes on such membranes.
Keywords
ESILDICBBTOFNBT2-DEPMSPVDFCoomassie Brilliant BlueMS/MSnitroblue tetrazoliumβ-nicotinamide adenine dinucleotidetwo-dimensional gel electrophoresisde novo sequencingpolyvinylidene difluorideRetinoic acidRetinal dehydrogenasetime of flightRetinalIdentificationTandem mass spectrometryphenazine methosulfateLaser desorption/ionizationNADelectrospray ionization
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Youji Shimazaki, Takahiro Kuroda,