Pyruvate Kinase

L’isoforme M2 du pyruvate kinase (PKM2) est responsable de la conversion du phosphoénolpyruvate (PEP) en pyruvate. Cette étape est la dernière étape de catalyse dans le processus de glycolyse puis joue un rôle essentiel dans la fermentation lactique effectuée chez les cellules anaérobiques, notamment les cellules myocardiques. De plus, le pyruvate kinase joue un rôle important dans le métabolisme des cellules cancéreuses. Au cours de cette expérience, nous caractérisent une mutante (K422R) de la PKM2 à l’aide de dosages enzymatiques des deux formes de l’enzyme en présence de certains effecteurs étant déjà identifiés comme régulateurs allostérique de la PKM2. La mutation a été confirmée à l’aide de séquençage et alignement de séquence avec la séquence protéique de la PKM2, retrouvée sur NCBI. Nos résultats démontrent que la mutation K422R favorise l’état T tétramérique et inactive de la PKM2 à cause de formation de liens stabilisantes au sein de la structure globulaire. La PKM2, sous sa forme active, existe plutôt sous l’état R tétramérique (3). Notre étude met en évidence une façon efficace de réguler l’activité enzymatique de la PKM2 à travers un changement dans la structure cristalline du type sauvage. Cette information s’avère utile pour des études dans le traitement du cancer qui vise à inhiber la fonction de la PKM2.

Carcinoma of the breast, the third most common cancer worldwide, accounts for the highest morbidity and mortality. Oxygen free radicals are able to cause dam- age to membranes, mitochondria and macromolecules including proteins, lipids and DNA. Accumulation of DNA damages has been suggested to contribute to carcinogenesis. In the present study, the relationship between oxidative stress and some enzyme activities in breast cancer was investigated. Malondialdehyde level, catalase, glu- tathione peroxidase, glucose-6-phosphate dehydrogenase, pyruvate kinase and arjinase activities in tumor tissues and adjacent normal tissues of 10 patients with invasive ductal carcinoma were determined. Higher malondialdehyde levels were observed in tumor tissues than noncancerous tissues (p<0.001). The activities of catalase, glutathione peroxidase, glucose-6- phosphate dehydrogenase, pyruvate kinase, arjinase in tumor tissues significantly increased compared to the control (p<0.001). We suggest...

To assess the effects of subordinate social status on digestive function, metabolism, and enzyme activity in salmonid fish, juvenile rainbow trout Oncorhynchus mykiss were paired with size-matched conspecifics (<1.5% difference in fork length) for 5 d. Fish that were fasted for 5 d and fish sampled directly from the holding tank were used as control groups. Both subordinate and fasted fish experienced significant decreases in intestine mass (P = 0.043), and the gall bladder showed marked and significant changes in both size (P = 0.004) and appearance. These findings suggest that the negative effect of social subordination on digestive function reflects in large part a lack of feeding. Hepatic phosphoenolpyruvate carboxykinase activity was significantly higher in subordinate fish relative to dominants, whereas subordinate hepatic pyruvate kinase activity was significantly lower; activities of both enzymes were significantly correlated with plasma cortisol concentrations and behavi...

The activity of pyruvate kinase present in a crude extract of the bloodstream form of Trypanosoma brucei was greatly increased by fructose 2,6-bisphosphate, which converted the saturation curve for phosphoenolpyruvate from a sigmoid into a hyperbola with no change in V. Phosphate and arsenate had an effect opposite to that of fructose 2,6-bisphosphate and the apparent Ka for fructose 2,6-bisphosphate was shifted from 75 nM to 1.5 microM by the presence of 5 mM phosphate. Fructose 1,6-bisphosphate had effects similar to those of fructose 2,6-bisphosphate but at approximately 4000-fold higher concentrations. Pyruvate kinases of Crithidia luciliae and of Leishmania major, two trypanosomatids which are like T. brucei in containing glycosomes, were also stimulated by fructose 2,6-bisphosphate and inhibited by phosphate.

Since chronic stress has been used widely for studying clinical depression and that brain energy metabolism and oxidative stress might be involved in the pathophysiology of this illness, the objective of this study was investigate the activities of pyruvate kinase, complex II and IV (cytocrome c oxidase) in hippocampus and prefrontal cortex of rats submitted to chronic variable stress. We also evaluated if vitamins E and C administration could prevent such effects. During 40 days adult rats from the stressed group were subjected to one stressor per day, at a different time each day, in order to minimize predictability. The stressed group had gained less weight while its immobilization time in the forced swimming test was greater than that of the control group. Results showed that stressed group presented an inhibition in the activities of complex II and cytochrome c oxidase in prefrontal cortex, while in hippocampus just complex IV was inhibited. Pyruvate kinase activity was not altered in stressed group when compared to control. Vitamins E and C administration prevented the alterations on respiratory chain caused by stress. These data suggest that the impairment of energy metabolism and oxidative stress could be related with the pathogenic pathways in stress related disorders.