Phosphoglucose isomerase and

For the assay of enzymes with products and reagents that have no absorption, fluorescence or luminescence in the ultraviolet or visible region, developments in analytical infrared spectroscopy can be used. In particular, mid-Fourier transform infrared (mFTIR) spectroscopy has been successfully applied to the determination of enzyme activities and kinetics, e.g. of /i-fructosidasc, phosphoglucose isomerase and polyphenol oxidase [90]. The method could very well be a tool that may also be applied to a variety of other enzyme classes. The potential of high-throughput applications, however, has yet to be demonstrated. 

An alternative method for detecting the fructose produced involves the use of ffuctokinase (EC 2.7.1.4), phosphoglucose isomerase, and glucose-6-phosphate dehydrogenase (EC 1.1.1.49), measuring in this case the nicotinamide-adenine dinucleotide phosphate, reduced form (NADPH) produced. ... 

Fructose 1,6-bisphosphate has attracted attention due to its important applications in the field of medicine, and is produced from glucose in three step by enzymatic reactions catalyzed by glucokinase, phosphoglucose isomerase, and phosphofructo-kinase. ATP is regenerated by acetate kinase (Fig. 17-26). Ishikawa and coworkers established an efficient method for production of fructose 1,6-bisphosphate in a... 

The effect on 2,3-BPG metabolism varies (Fig. 18.2). If the disorder is proximal to the 2,3-BPG shunt (e.g. deficiencies of hexokinase, phosphoglucose isomerase and aldolase A), the flow of metabolites through glycolysis will be decreased and consequently the concentration of 2,3-BPG will fall. If the deficiency is distal to the 2,3-BPG shunt (e.g. pyruvate kinase deficiency), the concentration of 2,3-BPG will rise. 

Roseman et al. (1958) purified the enzyme from microbial and mammalian extracts and demonstrated that fructose-6-phosphate, and not glucose-6-phosphate, was the required substrate. Ghosh et al. (1969) purified the enzyme from Neurospora crassa, E. coli, and rat liver until it was free from phosphoglucose isomerase and glutaminase. Using this more purified enzyme preparation, fructose-6-phosphate was shown to be the required substrate, and a 1 1 stoichiometry was demonstrated for glutamine and fructose-6-phosphate. 

Read, J., Pearce, J., Li, X., Muirhead, H., Chirgwin, I, and Davies, C. (2001) The crystal structure of human phosphoglucose isomerase at 1.6A resolution implications for catalytic mechanism, cytokine activity and haemolytic anaemia. J. Mol. Biol. 309, 447-463. 

Kruckeberg, A. L., Neuhaus, H. E., Feil, R.. Gottlieb, L. D., and Stitt, M. 1989. Decreased-activity mutants of phosphoglucose isomerase in the cytosol and chloroplast of Clarkia xantiana Impact on mass-action ratios and fluxes to sucrose and starch and estimation of flux control coefficients and elasticity coefficients. Biochem. J. 261, 457-467. 

FIGURE 9. (A) Crystal structure of the phosphoglucose isomerase (PGI) dimer from Pyrococcus furiosus (PDB ID 2GC2, Plate XVIII). (B) The Zn(II) active site of PGI and a bound phosphofrucose (2GC2, Plate XIX). (C) PGI with abound 5-phospho-D-arabinonohydroxamate inhibitor (2GC0, Plate XX). (D) PGI with a bound substrate mimic mannose 6-phosphate (2GC3, Plate XXI)... 

C. J. Jeffery, B.J. Bahnson, W. Chien, D. Ringe, and GA. Petsko. 2000. Crystal structure of rabbit phosphoglucose isomerase, a glycolytic enzyme that moonlights as neuroleukin, autocrine motility factor, and differentiation mediator... 

A fully carbamylated derivative of rabbit muscle phosphoglucose isomerase was utilized by James and Noltmann (1973), both for a quantitative amino-terminal analysis and for tryptic mapping experiments. Tryptic digestion of the derivative was restricted entirely to the arginyl residues. 

Cordeiro AT, Godoi PH, Delboni LE, Oliva G, Thiemann OH. Human phosphoglucose isomerase expression, purification, crystallization and preliminary crystallographic analysis. Acta Crystaliogr D Biol Crystaliogr 2001 57 592-5. 

Fig. 3. Phosphorylation of glucose and fructose. Hxkl Hexokinase PI Hxk2 hexokinase PII Glk glucokinase Pgi phosphoglucose isomerase Pfk phosphofructokinase Fbp fructose-1,6-bisphosphatase Pka protein kinase A...

Second, we selected or made several yeast glycolytical promoters including pyruvate kinase (Pyk) promoter, phosphoglucose isomerase (PCI) promoter, and phosphoglycerol kinase (PGK) promoter that can actively express genes in both E. coli and Saccharomyces yeasts and also use these promoters to express the E. coli genes both in yeast and E. coli. 

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