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Enzyme Activity Measurement for Pyruvate Kinase



Creative Enzymes is the most experienced service provider specialized in enzyme activity analysis. Utilizing state of the art technology and advanced equipment, Creative Enzymes is capable of handling all types of requests, offering the technical support and personalized service found nowhere else in the industry. Herein, Creative Enzymes is glad to provide the most accurate enzymatic assay for pyruvate kinase.

Pyruvate kinase (EC 2.7.1.40; ATP:pyruvate 2-O-phosphotransferase) catalyzes the final step of glycolysis, producing the second of the two ATP molecules generated in the glycolytic pathway. This enzyme catalyzes the irreversible phosphorylation of ADP at the expense of phosphoenolpyruvate (PEP), yielding pyruvate and ATP. This reaction is a key step leading to either anaerobic fermentation or oxidative phosphorylation of pyruvate, and it serves as a major regulator of glycolysis. Note that the allosteric regulation of this enzyme activity by various phosphorylated sugars serves as a vital role in maintaining an appropriate balance of intracellular substrate and product concentrations. Thus, the regulation of pyruvate kinase is crucial for controlling the levels of ATP, GTP, and glycolytic intermediates in the cell. Besides, pyruvate kinase can also act as a switch between the glycolytic and gluconeogenic pathways. Almost all pyruvate kinases exist as the homotetrameric protein with subunits of 50-60 kDa. Pyruvate kinase can be divided into four isozymes (L, R, M1 and M2) in mammals. Each isozyme performs different kinetic properties that report the particular metabolic requirements of the expressing tissue.

Pyruvate kinase plays a vital role in diagnostics, medicine, and drug development. For instance, genetic defects of pyruvate kinase cause the disease known as pyruvate kinase deficiency. Moreover, M2 pyruvate kinase in stool is a screening marker for colorectal cancer, and M2 pyruvate kinase in plasma is also a marker for follow-up studies during tumor therapy. M2 pyruvate kinase thus can also be a potentially attractive drug target. Therefore, quantification of the enzymatic activity of pyruvate kinase is critical to understanding its mechanisms and also sheds light on the design of potential new drugs for tumor therapy. Creative Enzymes has the capability of supplying the accurate activity assay available for pyruvate kinase. The pyruvate kinase reaction could be coupled to the lactate dehydrogenase reaction and assayed by monitoring the oxidation of NADH at 340 nm using spectrophotometry. Fully equipped with the most advanced spectrophotometric instruments, the leading technology, and a professional research team, the activity measurement for pyruvate kinase is assured in a reliable and timely manner. With years of discovery and development, Creative Enzymes has emerged as a worldwide leader in the field of enzymatic assays. Our high-quality enzyme assay services have been commended by more and more customers. Overall, Creative Enzymes has always been looking forward to the partnership with you in every step of research.

Figure: The  crystal structure of human M2 pyruvate kinase.
Figure: The crystal structure of human M2 pyruvate kinase.
PDB: 3SRH



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