Enzyme Activity Measurement for Glycerol Kinase

Creative Enzymes is an industrial biotech company specialized in enzyme activity assays. Our capabilities are extensive, covering enzyme assays in all steps of discovery, development, and optimization. As a reliable supplier, Creative Enzymes offers the services with high quality and competitive cost performance ratios. Herein, we are proud to provide the most precise activity measurement for glycerol kinase.

Glycerol kinase (EC 2.7.1.30; ATP:glycerol 3-phosphotransferase, GK) is an important metabolic enzyme at the interface of carbohydrate and lipid metabolism. GK catalyzes the rate-limiting step in glycerol utilization, the Mg-ATP dependent phosphorylation of glycerol which yields glycerol 3-phosphate (G3P). Apart from its biochemical functions, GK is also a “moonlighting” protein which functions as ATP-stimulated translocation promoter (ASTP). Moreover, GK has additional functions including binding to histones, interacting with porin, and playing a pivotal role in apoptosis. Furthermore, some studies have shown that GK could be involved in insulin sensitivity as it is overexpressed in response to thiazolidinediones, common drugs to treat type 2 diabetes mellitus. Lastly, the catalytic activity of GK is regulated at the protein level by inhibition by two allosteric effectors, the glucose-specific phospho-carrier protein of the phosphoenolpyruvate: sugar phosphotransferase system, III^Glc (also known as IIA^Glc), and the glycolytic intermediate, fructose 1,6- bisphosphate (FBP).

GK is ubiquitous in archaea, bacteria, and eukaryotes. To date, prokaryotic GKs have been the most widely studied. Of the eukaryotes, structural information is only available on Plasmodium falciparum GK. Thus, little is known about the structure and mechanism of GKs. One of the reasons for the limited amount of mechanistic information is the lack of a convenient activity assay. Fortunately, Creative Enzymes has developed and offers the accurate activity assay for GK. The enzyme activity of GK could be determined by using an ADP-coupled spectrophotometric assay. For example, the rate of conversion is monitored by coupling the reaction with the formation of lactate from pyruvate in the presence of NADH. The consumption rate of NADH is measured spectrophotometrically at 340 nm. Creative Enzymes is committed to being the most reliable supplier for enzymatic assays in the global market. We deliver the services in the shortest span of time from the date of order placement. The prompt service, best customer care, and dedicated resources have made Creative Enzymes the most preferred vendor.

Figure: The crystal structure of glycerol kinase from Trypanosoma brucei gambiense.
PDB: 5AZI