Mammalian electron transferring flavoprotein.flavoprotein dehydrogenase complexes observed by microelectrospray ionization-mass spectrometry and surface plasmon resonance

Microelectrospray ionization-mass spectrometry was used to directly observe electron transferring flavoprotein.flavoprotein dehydrogenase interactions. When electron transferring flavoprotein and porcine dimethylglycine dehydrogenase or sarcosine dehydrogenase were incubated together in the absence...

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Bibliographic Details
Published in:The Journal of biological chemistry 2004-04, Vol.279 (14), p.13786-13791
Main Authors: Hoard-Fruchey, Heidi M, Goetzman, Eric, Benson, Linda, Naylor, Stephen, Vockley, Jerry
Format: Article
Language:English
Subjects:
Acyl-CoA Dehydrogenase - chemistry
Acyl-CoA Dehydrogenase - metabolism
Animals
Binding, Competitive
Electron-Transferring Flavoproteins - chemistry
Electron-Transferring Flavoproteins - metabolism
Humans
Liver - enzymology
Mammals
Spectrometry, Mass, Electrospray Ionization - methods
Surface Plasmon Resonance - methods
Swine
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Summary:Microelectrospray ionization-mass spectrometry was used to directly observe electron transferring flavoprotein.flavoprotein dehydrogenase interactions. When electron transferring flavoprotein and porcine dimethylglycine dehydrogenase or sarcosine dehydrogenase were incubated together in the absence of substrate, a relative molecular mass corresponding to the flavoprotein.electron transferring flavoprotein complex was observed, providing the first direct observation of these mammalian complexes. When an acyl-CoA dehydrogenase family member, human short chain acyl-CoA dehydrogenase, was incubated with dimethylglycine dehydrogenase and electron transferring flavoprotein, the microelectrospray ionization-mass spectrometry signal for the dimethylglycine dehydrogenase.electron transferring flavoprotein complex decreased, indicating that the acyl-CoA dehydrogenases have the ability to compete with the dimethylglycine dehydrogenase/sarcosine dehydrogenase family for access to electron transferring flavoprotein. Surface plasmon resonance solution competition experiments revealed affinity constants of 2.0 and 5.0 microm for the dimethylglycine dehydrogenase-electron transferring flavoprotein and short chain acyl-CoA dehydrogenase-electron transferring flavoprotein interactions, respectively, suggesting the same or closely overlapping binding motif(s) on electron transferring flavoprotein for dehydrogenase interaction.
ISSN:0021-9258