A NADH-dependent transketolase assay in erythrocyte hemolysates
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Methods for assessment of thiamine (vitamin B<inf>1</inf>)
2018, Laboratory Assessment of Vitamin StatusThe uremic toxin oxythiamine causes functional thiamine deficiency in end-stage renal disease by inhibiting transketolase activity
2016, Kidney InternationalCitation Excerpt :This may have contributed to discordant reports of decreased transketolase activity in ESRD patients. Studies by Lonergan et al.2,3 and Kopple et al.41 used a modified method of transketolase activity assessment developed by Dreyfus,42 which was subsequently found to lack specificity and reproducibility43–45 and is no longer used. Inhibition of transketolase would have been lost if high dilution of red blood cell lysate was prepared without immediate assay of transketolase activity such that OTPP dissociated from the holoenzyme.
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2014, Seminars in Pediatric NeurologyCarbon metabolism and the sign of control coefficients in metabolic adaptations underlying K-ras transformation
2011, Biochimica et Biophysica Acta - BioenergeticsCitation Excerpt :Reactions were initiated by the addition of glucose to a final concentration of 10 mM. Transketolase (TKT, EC 2.2.1.1) activity was determined using the enzyme linked method described by Smeets and colleagues [32]. Briefly, samples were added to a cuvette containing 5 mM MgCl2, 0.2 U/ml triose phosphate isomerase, 0.2 mM NADH, 0.1 mM thiamine pyrophosphate in 50 mM Tris–HCl, and pH 7.6 at 37 °C.
Gene silencing of TKTL1 by RNAi inhibits cell proliferation in human hepatoma cells
2007, Cancer LettersCitation Excerpt :The resulting supernatant was filtered to remove some endogenous metabolites. TK activity was determined by using enzyme-linked method [13]. Samples were added to a cuvette containing buffer (50 mM Tris/HCl, pH 7.6), 2 mM ribose 5-phosphate, 1 mM xylulose 5-phosphate, 5 mM MgCl2, 0.2 U mL−1 of TPI, 0.2 mM NADH and 0.1 mM TPP.
Protocol and application of basal erythrocyte transketolase activity to improve assessment of thiamine status
2023, Annals of the New York Academy of Sciences