PMID: 9426198Jan 13, 1998Paper

Medium-chain acyl CoA dehydrogenase: evidence for phosphorylation

Biological Chemistry
Peter MacherouxSandro Ghisla

Abstract

Mature medium chain acyl-CoA dehydrogenase isolated from pig kidney (pkMCADH) and originating from mitochondria carries a phosphate group as demonstrated by 31P-NMR-spectroscopy and chemical analysis. Two broad resonances at -6.3 and -8 ppm are observed and are assigned to the pyrophosphate group of the cofactor FAD. A third, narrow resonance at 4.65 ppm indicates the presence of a phosphomonoester residue. Chemical analysis of intact pkMCADH shows the presence of 3 +/- 0.3 phosphates, those of FAD and of an additional covalently attached phosphate. With recombinant, human wild type MCADH expressed in and purified from E. coli only the two FAD phosphates (2 +/- 0.35) are found. Similarly, pkMCADH which has been converted to the apoenzyme and reconstituted to holoenzyme also contains 2 +/- 0.4 phosphates. The covalently bound phosphate can be hydrolyzed by phosphatase and subsequently removed by dialysis. The phosphate group has no detectable effect on the catalytic activity of the MCADH measured with artificial and natural electron acceptors such as pig electron transferring flavoprotein. However, phosphorylation has a marked effect on protein solubility which is +5-fold lower for the dephosphorylated protein.

References

Nov 15, 1979·Analytical Biochemistry·P A LanzettaO A Candia
Apr 15, 1989·European Journal of Biochemistry·S Ghisla, V Massey
Sep 1, 1981·Analytical Biochemistry·E H Mayer, C Thorpe
Jun 1, 1995·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·C Thorpe, J J Kim
Aug 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·J J KimR Paschke

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Citations

Nov 13, 2012·Molecular Genetics and Metabolism·Kaitlyn KormanikAl-Walid Mohsen
Feb 18, 2014·Acta Crystallographica. Section D, Biological Crystallography·Jian YuMin Yao
Apr 11, 2013·Physiological Genomics·Dillon J ChungJames F Staples

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