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Deficiency of AdoCbl results in decreased MMCoA mutase activity, accumulation of methylmalonyl-CoA, and subsequent production of methylmalonic acid.
2] Nonstandard abbreviations: MS/MS, tandem mass spectrometry; MMA, methylmalonic acidemia; MMCoA, methylmalonyl-CoA; AdoCbl, 5-deoxy adenosylcobalamin.
Among different Cbl species, only MeCbl and AdoCbl are metabolically active, functioning as cofactors for the cytoplasmic enzyme MS and the mitochondrial enzyme methylmalonyl-CoA mutase, respectively.
Treating SH-SY5Y cells with 1nM NRG-1 for one hour resulted in significantly increased levels of metabolically active AdoCbl and MeCbl, while decreasing the level of CNCbl without changing the level of S[O.
To test our hypothesis that NRG-1 promoted the formation of AdoCbl and MeCbl by increasing the GSH level, we first determined whether synthesis of active Cbl species was GSH-dependent.
The decrease in CNCbl caused by NRG-1, along with increased AdoCbl and MeCbl, suggests that NRG-1 promotes the conversion of inactive CNCbl to active AdoCbl and MeCbl (Figures 1(e)1(g)).
Standard solutions containing 100 pmol/L OHCbl, CNCbl, AdoCbl, and MeCbl (all from Sigma-Aldrich) were separated with HPLC to determine the retention times of the different Cbls.
Previous studies suggested that TC carries primarily OHCbl and to some degree AdoCbl (18).
4] Nonstandard abbreviations: Cbl, cobalamin; TC, transcobalamin; CNCbl, cyanocobalamin; OHCbl, hydroxocobalamin; AdoCbl, 5'-deoxyadenosylcobalamin; MeCbl, methylcobalamin; TC-CNCbl, TC-bound CNCbl.
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