Flavinylation of the precursor of mitochondrial dimethylglycine dehydrogenase by intact and solubilised mitochondria.
The flavinylation and the presequence processing of the mitochondrial matrix enzyme dimethylglycine dehydrogenase (Me(2)GlyDH) were investigated with the reticulocyte lysate translated precursor (pMe(2)GlyDH) added to solubilised mitoplasts of rat liver mitochondria. The flavinylation of pMe(2)GlyDH was strictly dependent on the addition of mitochondrial protein(s), among which the mitochondrial flavinylation stimulating factor [Brizio C., et al. (2000) Eur. J. Biochem 267, 4346-4354], that actively promotes holo-Me(2)GlyDH formation. The precursor processing, that accompanies the biogenesis of the enzyme, was not required to allow the flavinylation to proceed. The comparison of the time course of the flavinylation and the processing of pMe(2)GlyDH demonstrated that the covalent attachment of the flavin moiety preceded the presequence processing by mitochondrial processing peptidase.
Brizio
Carmen
C
Dipartimento di Biochimica e Biologia Molecolare, Università di Bari, C.N.R., Via Orabona 4, Italy.
Barile
Maria
M
Brandsch
Roderich
R
D000818:Animals; D050893:Dimethylglycine Dehydrogenase; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000378:metabolism*; D005415:Flavins / Q000378:metabolism*; D020035:Holoenzymes / Q000096:biosynthesis; D007700:Kinetics; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology*; D024101:Mitochondrial Proteins; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012995:Solubility; D014357:Trypsin / Q000378:metabolism
D004792:Enzyme Precursors; D005415:Flavins; D020035:Holoenzymes; D024101:Mitochondrial Proteins; D010089:Oxidoreductases, N-Demethylating; D050893:Dimethylglycine Dehydrogenase; C494400:Dmgdh protein, rat; D014357:Trypsin