Novel and simple high-performance liquid chromatographic method for determination of 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity.
We present here a high-performance liquid chromatographic method for the evaluation of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity. The automated method was applied to fungal and mouse liver extracts and validated by the addition of mevastatin to the reaction mixture and by several intra- and inter-day assays. This method offers important advantages over those previously reported because no radiolabeled substrates or expensive techniques such as mass spectrometry are required, and the time of analysis is relatively short. Moreover, the method can be successfully applied to different biological samples; hence, it should be very useful in evaluating potential inhibitors of the HMG-CoA enzyme and investigating cholesterol metabolism, cell growth and differentiation processes.
Buffalini
Michele
M
Istituto di Chimica Biologica Giorgio Fornaini, Università degli Studi di Urbino Carlo Bo, via A. Saffi 2, 61029 Urbino (PU), Italy.
Pierleoni
Raffaella
R
Guidi
Chiara
C
Ceccaroli
Paola
P
Saltarelli
Roberta
R
Vallorani
Luciana
L
Zeppa
Sabrina
S
Stocchi
Vilberto
V
D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005658:Fungi / Q000201:enzymology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D051379:Mice; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D006903:Hydroxymethylglutaryl CoA Reductases