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Moreover our results indicate that
Moreover, our results indicate that ADP is by far a better phosphate donor than AMP (Table 2). Yet, the bindings of AMP and ATP are mutually exclusive, whereas the bindings of ADP and ATP are additive (Fig. 7). The multiplicity of the phosphate donors in the TgAK reaction (Tables 1 & 2), suggests that all the phosphate donors bind to a single site. In consequence, it Pentamidine dihydrochloride sale is plausible that additivity results from a reaction with ADP as depicted above. Notwithstanding ATP binding before Ado or vice versa, it is possible that the release of ADP from the ATP site is rate limiting, i.e., without this release no further catalysis by ATP is possible. Indeed, the rate of an enzymic reaction is governed by binding of substrate and/or release of product. Hence, the binding of Ado may hinder the binding of ATP and vice versa (Fig. 2). But the efficiency of TgAK attests that this effect is likely a result of conformational changes associated with the ligand binding process, which in no way hinders catalysis. Our results also indicate that AMP is readily releasable from the Ado site as its Kii is 0.5mM; therefore, the release of ADP must be the key determinant of further catalysis. The fact that we could not obtain any inhibitory pattern with ADP as product inhibitor of the TgAK reaction with varied ATP, which indicates that the bindings of ATP and ADP are not mutually exclusive (Fig. 7) as both ATP and ADP bind to a single site (Zhang et al., 2006), is a sufficient proof for this proposition.
In conclusion, the mechanism of TgAK is a hybrid random bi-uni ping-pong uni-bi as depicted in the above scheme. Thus, the overall reaction of TgAK could be portrayed as the sum of two partial reactions in which enzyme bound ADP acts as an intermediate phosphate donor as follows:
A steady-state treatment of the proposed mechanism is too complex to make possible derivation of a rate equation. An equation based on rapid equilibrium, using the method of Cha (1968) would have been quite manageable. A requisite of rapid equilibrium, however, is that the binding of one ligand has no effect on the binding of another. Our initial velocity studies clearly indicate that the binding of Ado decreases the binding of ATP and vice versa, which precludes resorting to rapid equilibrium. Derivation of rate equation for this reaction must hence await the development of computer programs that could solve such steady state equation.
Introduction
Diabetes, especially type 2 diabetes, is now a global epidemic that reflects the harmful consequences of modern lifestyles. Consumption of processed, high-calorie foods, along with physical inactivity, has created an imbalance between energy intake and expenditure [1,2]. This disruption of energy balance is characterized by shifts in lipid and glucose metabolism manifesting as fasting and postprandial hyperglycemia together with dyslipidemia and is further promoted by insulin resistance in a number of tissues [3]. Chronic exposure to glucose overload, free fatty acids, and amino acids can be toxic [4]. Chronic kidney disease (CKD) associated with diabetes is a relatively common manifestation, with both the prevalence and incidence rising in recent years [5]. CKD is the end result of multifactorial processes that mostly stem from deranged glucose and lipid metabolism. This is demonstrated by the increased expression of proteins in relevant signaling pathways and consistent pathologic renal findings. Many studies and experiments have been devised to elucidate the pathogenesis of these metabolic disruptions, although emerging concepts related to such findings remain vast and vague. In an attempt to unravel the core of this disease entity, a growing body of evidence shows that dysregulation of 5′ adenosine monophosphate–activated protein kinase (AMPK) in relevant tissues is crucial to the development of metabolic syndrome and diabetes [6]. Therefore, targeting this enzyme is a matter of great interest, as it may ameliorate some of the pathologic features of the disease. AMPK regulates the coordination of anabolic processes with its activation proven to improve glucose and lipid homeostasis in insulin-resistant animal models [7,8]. AMPK is strongly expressed in the kidney, where it is involved in diverse physiological and pathologic processes, including ion transport, podocyte function, and diabetic renal hypertrophy. In light of this, we have looked for novel agents that would help to modulate AMPK in an organ-specific manner that targets the diabetic kidney. Instead of providing an exhaustive review of the role of AMPK in these pathologies, this review aims to discuss the mechanisms of AMPK agonists in the context of diabetic nephropathy (DN) and, therefore, shed light on the risks and benefits of currently available and promising future AMPK activators as a treatment option.