64060-84-0Relevant academic research and scientific papers
BOROHYDRIDE REDUCTION OF PERIODATE-OXIDIZED NUCLEOTIDES; ISOLATION AND STRUCTURE OF THE REDUCTION INTERMEDIATE
Rosenthal, Luann P.,Hogenkamp, Harry P. C.,Bodley, James W.
, p. 85 - 92 (1982)
The reduction of periodate-oxidized nucleotides with sodium borohydride proceeds via a reaction intermediate presumed to be a monoalcohol.The borohydride-reduction intermediate of periodate-oxidized ADP has been isolated by anionexchange, liquid chromatography, and subjected to further reduction.Using sodium borohydride and sodium borodeuteride alternately in the two reduction steps, it was determined, by 1H-n.m.r.-spectral analysis, that the two aldehyde groups are sequentially reduced in the order 3' and 2', and it was concluded that the isolated intermediate corresponds to the semi-reduced, 3'-alcohol, 2'-aldehyde derivative.This compound should be a useful analog for the study of enzymes and proteins that interact with nucleotides.
Inactivation of lactobacillus leichmannii ribonucleotide reductase by 2',2'-difluoro2'-deoxycytidine s'-triphosphate: Covalent modification
Lohman, Gregory J.S.,Stubbe, Joanne
experimental part, p. 1404 - 1417 (2011/02/21)
Ribonucleotide reductase (RNR) from Lactobacillus leichmannii, a 76 kDa monomer using adenosylcobalamin (AdoCbl) as a cofactor, catalyzes the conversion of nucleoside triphosphates to deoxynucleotides and is rapidly ( 3H]- and [5-3H]F2CTP were synthesized and used independently to inactivate RNR. Sephadex G-50 chromatography of the inactivation mixture revealed that 0.47 equiv of a sugar was covalently bound to RNR and that 0.71 equiv of cytosine was released. Alternatively, analysis of the inactivated RNR by SDS-PAGE without boiling resulted in 33% of RNR migrating as a 110 kDa protein. Inactivation of RNR with a mixture of [1'-3H]F2CTP and [1'-2H]F 2CTP followed by reduction with NaBH4, alkylation with iodoacetamide, trypsin digestion, and HPLC separation of the resulting peptides allowed isolation and identification by MALDI-TOF mass spectrometry (MS) of a 3H/2H-labeled peptide containing C731 and C736 from the C-terminus of RNR accounting for 10% of the labeled protein. The MS analysis also revealed that the two cysteines were cross-linked to a furanone species derived from the sugar of F2CTP. Incubation of [1-3H]F2CTP with C119S-RNR resulted in 0.3 equiv of sugar being covalently bound to the protein, and incubation with NaBH4 subsequent to inactivation resulted in trapping of 2'-fluoro-2'-deoxycytidine. These studies and the ones in the preceding paper (DOI: 10.1021/bi9021318) allow proposal of a mechanism of inactivation of RNR by F2CTP involving multiple reaction pathways. The proposed mechanisms share many common features with F2CDP inactivation of the class I RNRs.
