The Thionicotinamide Analogs of DPN and TPN. I. Preparation and Analysis^*

Abstract

The synthesis of the thionicotinamide analog of triphosphopyridine nucleotide catalyzed by pig brain transglycosidase is described. The thionicotinamide analogs of both diphospho-and triphosphopyridine nucleotide have been purified from the corresponding nicotinamide nucleotides by preparative gradient elution chromatography, and characterized by elementary analyses. The spectrophotometric constants reported are based on ribose, phosphate, and adenine analyses of the samples.

Suppression of Cytosolic NADPH Pool by Thionicotinamide Increases Oxidative Stress and Synergizes with Chemotherapy

“…Previous studies have shown that NADS and NADPS can be synthesized from thionicotinamide using porcine liver powder (Stein et al, 1963). Using an enzymatic assay, NADS can be phosphorylated to NADPS by NADK.…”

Site-Directed Mutagenesis of the Redox-Active Cysteines of Trypanosoma cruzi Trypanothione Reductase

“…Cells were harvested by centrifugation (6000 g, 20 rnin), washed free of medium with 0.9 % (masshol.) NADPH stock solutions were determined spectrophotometrically at 340 nm using an absorption coefficient of 6.22X103 M-' cm-' [25] ; the thio-NADP' concentration was determined at 260 nm using an absorption coefficient of 1.95X104 M-' cm-' [26]. Cells were lysed by repeated sonication and, following centrifugation (I4 000 g, 30 rnin), the resulting cell-free extract was adjusted to 0.4 % (mass/vol.)…”

Structural and Functional Characterization of Plant Aminoaldehyde Dehydrogenase from Pisum sativum with a Broad Specificity for Natural and Synthetic Aminoaldehydes

“…For calculations, absorption maxima and extinction coefficients of reduced forms of NAD + analogs were taken from the literature. 26,27 The rate of NAD + -mediated reaction was arbitrarily taken as 100%. CPAL, 3-cyanopropionaldehyde; DMABAL, N,Ndimethyl-4-aminobutyraldehyde.…”

“…6 NAD + analogs were assayed as coenzymes at a concentration of 500 μM with 1 mM APAL as a substrate. The following absorption maxima and extinction coefficients of reduced forms of these analogs 26,27 were used for calculations: NADPH, ɛ 340 = 6.62 mM − 1 cm − 1 ; thio-NADH, ɛ 400 = 11.9 mM − 1 cm − 1 ; 3-pyridinealdehyde-NADH, ɛ 358 = 9.3 mM − 1 cm − 1 , 3-acetylpyridine-NADH, ɛ 363 = 9.1 mM − 1 cm − 1 ; deamino NADH, ɛ 338 = 6.2 mM − 1 cm − 1 . Tris-HCl buffers (150 mM) in the pH range of 7.0-9.0 and glycine-NaOH buffers (150 mM) in the pH range of 8.8-10.6 were used to determine pH optimum using 1 mM APAL and 0.5 mM NAD + .…”