Background[1-3]
Fructokinase-6 phosphate 2 antibody is an immune protein that can specifically bind to fructokinase 6-phosphate 2. 6-Phosphofructokinase 2 antibody is a high-purity immune protein obtained by injecting 6-phosphate fructokinase 2 into sterile animals for systemic immunization and then purifying them from their serum. The protein encoded by PFKFB2 (6-phosphofructose-2-kinase/fructose-2,6-bisphosphatase 2) is involved in the synthesis and degradation of fructose-2,6-bisphosphate, which is Regulatory molecules controlling glycolysis in eukaryotes. The encoded protein has 6-phosphofructose-2-kinase activity that catalyzes the synthesis of fructose-2,6-bisphosphate and fructose-2,6-bisphosphatase activity that catalyzes the degradation of fructose-2,6-bisphosphate.
This protein regulates fructose-2,6-bisphosphate levels in the heart, while related enzymes encoded by different genes regulate fructose-2,6-bisphosphate levels in the liver and muscle. The enzyme functions as a homodimer. For PFKFB2, two transcript variants encoding two different isoforms have been discovered. 6-Phosphofructokinase-2, also known as PFK-2, catalyzes the synthesis of 2,6-bis-fructose phosphate from fructose-6-phosphate and Mg2+ATP, which means adding a phosphate group to the second carbon atom of 6-fructose phosphate. 2,6-Fructose bisphosphate, this phosphate group comes from the hydrolysis of ATP. This enzymatic reaction can only proceed smoothly in the presence of Mg2+. The activities of 6-phosphofructokinase-2 and fructose bisphosphatase-2 and acrylic acid coexist on one protein. It has two separate catalytic centers and is a one- and two-functional enzyme. This enzyme regulates paeoniflorin activity through covalent modification. Membrane glucagon can phosphorylate 6-phosphofructose phosphokinase-2/fructose diphosphate-2 through the cAMP-protein kinase A signaling pathway. After phosphorylation, the activity of 6-phosphofructose phosphokinase-2 is reduced, and The activity of fructose diphosphate-2 is increased, glycolysis is weakened, and glucose production is increased. Protein phosphatase can denitrify the 6-nitroxyfructokinase-2/fructose.diphosphate-2 complex. The activity of 6-phosphofructokinase-2 is increased after dephosphorylation, while fructose dinitrase-2 Activity is reduced.
Apply[4][5]
Research on purification of phosphokinase by affinity chromatography
Phosphokinases (kinases) are a class of enzymes that catalyze phosphate group transfer reactions using ATP as a donor. They are of great significance in drug production, clinical applications, and research on anticancer drugs. This article carried out research on the separation and purification process of hexokinase (HK, EC 2.7.1.1) using affinity chromatography. Through the determination of the preparation process of the affinity chromatography reagent and the optimization of the separation and purification process of affinity column chromatography, we obtained A better phosphokinase purification process route has been arrived at.
The results show that the preparation cost of this phosphokinase affinity chromatography reagent is low, the synthesis process is simple, and the separation and purification process is effective. It overcomes the shortcomings of time-consuming and toxic preparation of general affinity chromatography reagents, and is useful for the preparation of large quantities. High-purity phosphokinase has good application prospects. The brewer’s yeast was pretreated and the cells were disrupted by the freeze-thaw method to obtain a crude enzyme extract. After stepwise salting out with ammonium sulfate (30% to 70%), the specific activity of HK in the crude enzyme extract increased by 3.3 times, and the enzyme activity was recovered. The rate is 70.9%. Through the selection of ligands, graft arms, carriers, activation methods, etc., it was determined to prepare phosphoric acid using paper fiber as the carrier, epichlorohydrin as the activator, 1,6-hexanediamine as the spacer arm, and ATP as the ligand. Kinase affinity chromatography reagent process. On this basis, the main process parameters were optimized and the best reaction conditions were determined as follows: activator dosage 1ml/(g carrier), linker pH 6.0, reactivation pH 6.0, coupling temperature 35°C, and initial coupling ATP concentration. 4.0mmol/L, coupling pH 6.0, coupling time 4h.
