scholarly journals Isolation and properties of the glycolytic enzymes from Zymomonas mobilis. The five enzymes from glyceraldehyde-3-phosphate dehydrogenase through to pyruvate kinase

1986 ◽  
Vol 238 (1) ◽  
pp. 275-281 ◽  
Author(s):  
A Pawluk ◽  
R K Scopes ◽  
K Griffiths-Smith
Keyword(s):  
Pyruvate Kinase ◽  
Zymomonas Mobilis ◽  
Specific Activity ◽  
High Specific Activity ◽  
Phosphoglycerate Kinase ◽  
Glycolytic Enzymes ◽  
Phosphoglycerate Mutase ◽  
Eukaryotic Species ◽  
Sugar Phosphates ◽  
Subunit Size

The five glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, enolase and pyruvate kinase were each purified from extracts of Zymomonas mobilis cells, by using dye-ligand chromatography as the principal step. Two procedures, producing three and two of the enzymes respectively, are described in detail. Z. mobilis glyceraldehyde-phosphate dehydrogenase was found to be similar in most respects to the enzyme from other sources, except for having a slightly larger subunit size. Phosphoglycerate kinase has properties typical for this enzyme; however, it did not show the sulphate activation effects characteristic of this enzyme from most other sources. Phosphoglycerate mutase is a dimer, partially independent of 2,3-bisphosphoglycerate, and has a high specific activity. Enolase was found to be octameric; otherwise its properties were very similar to those of the yeast enzyme. Pyruvate kinase is unusual in being dimeric, and not requiring K+ for activity. It is not allosterically activated by sugar phosphates, having a high activity in the absence of any effectors. Some quantitative differences in the relative amounts of these enzymes, compared with eukaryotic species, are ascribed to the fact that Z. mobilis utilizes the Entner-Doudoroff pathway rather than the more common Embden-Meyerhoff glycolytic route.

scholarly journals Inhibition of glycolytic enzymes by 2-phosphotartronate

1976 ◽  
Vol 153 (3) ◽  
pp. 741-744 ◽  
Author(s):  
M K Thomas ◽  
T G Spring
Keyword(s):  
Pyruvate Kinase ◽  
Competitive Inhibition ◽  
Phosphoglycerate Kinase ◽  
Triose Phosphate Isomerase ◽  
Glycolytic Enzymes ◽  
Phosphoglycerate Mutase ◽  
Rabbit Muscle ◽  
Triose Phosphate ◽  
Permanganate Oxidation

2-Phosphotartronate has been synthesized by permanganate oxidation of glycerol 2-phosphate and has been tested as an inhibitor of five glycolytic enzymes that bind phosphoglycerate or phosphoglycollate. Competitive inhibition of rabbit muscle phosphoglycerate mutase, enolase and pyruvate kinase was observed. Triose phosphate isomerase and 3-phosphoglycerate kinase were not inhibited.

scholarly journals Activity and androgenic control of glycolytic enzymes in the epididymis and epididymal spermatozoa of the rat

1976 ◽  
Vol 156 (3) ◽  
pp. 527-537 ◽  
Author(s):  
D E Brooks
Keyword(s):  
Phosphoglycerate Kinase ◽  
Glycolytic Enzymes ◽  
Phosphoglycerate Mutase ◽  
Enzyme Release ◽  
Glycerol Phosphate ◽  
Phosphogluconate Dehydrogenase ◽  
Duct Ligation ◽  
Epididymal Spermatozoa ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Glycerol Phosphate Dehydrogenase

1. Procedures were developed for the extraction and assay of glycolytic enzymes from the epididymis and epididymal spermatozoa of the rat. 2. The epididymis was separated into four segments for analysis. When rendered free of spermatozoa by efferent duct ligation, regional differences in enzyme activity were apparent. Phosphofructokinase, glycerol phosphate dehydrogenase and glucose 6-phosphate dehydrogenase were more active in the proximal regions of the epididymis, whereas hexokinase, lactate dehydrogenase and phosphorylase were more active in the distal segment. These enzymes were less active in the epididymis of castrated animals and less difference was apparent between the proximal and distal segments. However, the corpus epididymidis from castrated rats had lower activities of almost all enzymes compared with other epididymal segments. 3. Spermatozoa required sonication to obtain satisfactory enzyme release. Glycolytic enzymes were more active in spermatozoa than in epididymal tissue, being more than 10 times as active in the case of hexokinase, phosphoglycerate kinase and phosphoglycerate mutase. 4. The specific activities of a number of enzymes in the epididymis were dependent on the androgen status of the animal. These included hexokinase, phosphofructokinase, aldolase, glyceraldehyde phosphate dehydrogenase, phosphoglycerate kinase, pyruvate kinase, glycerol phosphate dehydrogenase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and phosphorylase. 5. The caput and cauda epididymidis differed in the extent to which enzyme activities changed in response to an altered androgen status. The most notable examples were hexokinase, phosphofructokinase, aldolase, phosphoglycerate kinase, 6-phosphogluconate dehydrogenase and phosphorylase.

Activities and Regulation of Enzymes of Carbohydrate Metabolism in Spruce ( Picea abies)

1991 ◽  
Vol 46 (7-8) ◽  
pp. 597-604 ◽  
Author(s):  
Meinrad Boll
Keyword(s):  
Picea Abies ◽  
Cell Suspension ◽  
Specific Activity ◽  
Phosphoglycerate Kinase ◽  
Cell Suspension Cultures ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Glycolytic Enzymes ◽  
Strong Increase ◽  
Fructose 1,6 Bisphosphate

Abstract Activities of the glycolytic enzymes were determined in seedlings, callus cultures and cell sus­ pension cultures of spruce (Picea abies) (L.) (Karst). The rate-limiting enzymes of the pathway were the hexokinases, ATP: phosphofructo-kinase, fructose-1,6-bisphosphatase and pyruvate kinase. Two phosphofructokinases were found: ATP : fructose-6-phosphate 1-phosphotransferase (PFK) and pyrophosphate :fructose-6-phosphate 1-phosphotransferase (PFP). In the presence of its activator fructose-2,6-bisphos-phate, PFP had a 4 -5-fold higher specific activity than PFK. PFP could be activated about 20-fold by fructose-2,6-bisphosphate at saturating concentrations of the substrates (fructose-6-phosphate and pyrophosphate). The increase of Vmax was accompanied by a strong increase in the apparent affinity of the enzyme for the substrates. Km for fructose-6-phosphate and pyrophosphate was 0.44 mM and 24 μM, respectively. Ka for fructose-2,6-bisphosphate was 24 nM. In seedlings, specific activity of the glycolytic enzymes was 30-300 percent higher in the hypocotyls, except for fructose-1,6-bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase, their activity being 100-150percent higher in the cotyledons, This distribution remained unchanged during periods of 2 -16 weeks of cultivation of the seedlings. In callus cultures and in cell suspension cultures, grown mixotrophically with different car­ bohydrates, all enzymes were between 1-and 7-fold higher than in autotrophically grown seed­ lings. Incubation of seedlings in mineral salt mixture containing a carbohydrate resulted in a rapid coordinate increase of the activities to the levels of callus-or cell suspension cultures. This induction required a carbohydrate and oxygen. During prolonged cultivation of cell suspension cultures, when carbohydrate became limiting, activity of the enzymes slowly declined.

scholarly journals A moonlighting role for enzymes of glycolysis in the co-localization of mitochondria and chloroplasts

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Youjun Zhang ◽  
Arun Sampathkumar ◽  
Sandra Mae-Lin Kerber ◽  
Corné Swart ◽  
Carsten Hille ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Pyruvate Kinase ◽  
Protein Interactions ◽  
Glycolytic Enzymes ◽  
Current Understanding ◽  
Phosphoglycerate Mutase ◽  
Protein Protein Interactions ◽  
Photosynthetic Eukaryotes ◽  
Substrate Channelling ◽  
Knockout Mutation

Abstract Glycolysis is one of the primordial pathways of metabolism, playing a pivotal role in energy metabolism and biosynthesis. Glycolytic enzymes are known to form transient multi-enzyme assemblies. Here we examine the wider protein-protein interactions of plant glycolytic enzymes and reveal a moonlighting role for specific glycolytic enzymes in mediating the co-localization of mitochondria and chloroplasts. Knockout mutation of phosphoglycerate mutase or enolase resulted in a significantly reduced association of the two organelles. We provide evidence that phosphoglycerate mutase and enolase form a substrate-channelling metabolon which is part of a larger complex of proteins including pyruvate kinase. These results alongside a range of genetic complementation experiments are discussed in the context of our current understanding of chloroplast-mitochondrial interactions within photosynthetic eukaryotes.

scholarly journals The effect of dietary and hormonal conditions on the activities of glycolytic enzymes in rat epididymal adipose tissue

1969 ◽  
Vol 115 (3) ◽  
pp. 405-417 ◽  
Author(s):  
E. D. Saggerson ◽  
A. L. Greenbaum
Keyword(s):  
Adipose Tissue ◽  
Lactate Dehydrogenase ◽  
Pyruvate Kinase ◽  
High Fat Diet ◽  
Alloxan Diabetes ◽  
Phosphoglycerate Kinase ◽  
Glycolytic Enzymes ◽  
Glucose Phosphate Isomerase ◽  
High Fat ◽  
Glucose Phosphate

1. Measurements were made of the activities of nine glycolytic enzymes in epididymal adipose tissues obtained from rats that had undergone one of the following treatments: starvation; starvation followed by re-feeding with bread or high-fat diet; feeding with fat without preliminary starvation; alloxan-diabetes; alloxan-diabetes followed by insulin therapy. 2. In general, the activities of the glycolytic enzymes of adipose tissue, unlike those of liver, were not greatly affected by the above treatments. 3. The ‘key’ glycolytic enzymes, phosphofructokinase and pyruvate kinase, were generally no more adaptive in response to physiological factors than other glycolytic enzymes such as glucose phosphate isomerase, fructose diphosphate aldolase, triose phosphate isomerase, glycerol 3-phosphate dehydrogenase, phosphoglycerate kinase and lactate dehydrogenase. 4. Adiposetissue pyruvate kinase did not respond to feeding with fat in a manner similar to the liver enzyme. 5. Glyceraldehyde phosphate dehydrogenase had a behaviour pattern unlike the other eight glycolytic enzymes studied in that its activity was depressed by feeding with fat and was not restored to normal by re-feeding with a high-fat diet after starvation. These results are discussed in relation to the requirements of adipose tissue for glycerol phosphate in the esterification of fatty acids. 6. A statistical analysis of the results permitted the writing of linear equations describing the relationships between the activities of eight of the enzymes studied. 7. Evidence is presented for the existence of two constant-proportion groups amongst the enzymes studied, namely (i) glucose phosphate isomerase, phosphoglycerate kinase and lactate dehydrogenase, and (ii) triose phosphate isomerase, fructose diphosphate aldolase and pyruvate kinase. 8. Mechanisms for maintaining the observed relationships between the activities of the enzymes in the tissue are discussed.

scholarly journals Induction of glycolytic enzyme synthesis in proliferating fibroblasts. Study of phosphofructokinase, glucose phosphate isomerase and pyruvate kinase

1983 ◽  
Vol 214 (1) ◽  
pp. 195-201 ◽  
Author(s):  
M C Meienhofer ◽  
J C Dreyfus ◽  
A Kahn
Keyword(s):  
Enzyme Activity ◽  
Pyruvate Kinase ◽  
Specific Activity ◽  
Human Fibroblasts ◽  
Glycolytic Enzymes ◽  
Resting Cells ◽  
Glucose Phosphate Isomerase ◽  
Antigen Concentration ◽  
Quiescent Cells ◽  
Glucose Phosphate

Specific activity of phosphofructokinase is 7-8-fold higher in exponentially growing human fibroblasts than in quiescent cells, but the difference is considerably less pronounced for two other glycolytic enzymes, glucose phosphate isomerase and pyruvate kinase. The ratio of the F-type to L-type phosphofructokinase subunits is essentially the same in growing and resting cells, 4:1. F-type-phosphofructokinase-related antigen concentration is decreased in resting cells as compared with proliferating fibroblasts, but relatively less than the enzyme activity; the ratio of the enzyme activity to the antigen concentration (immunological specific activity) is therefore lower in resting than in growing fibroblasts. Synthesis of phosphofructokinase, as a percentage of the total protein synthesis, is about 30-fold greater during the proliferative phase than in quiescent cells, but this difference is only 3-4-fold for glucose phosphate isomerase and pyruvate kinase. Modulation of the synthesis of phosphofructokinase therefore seems to be responsible for the changes of its specific activity in function of cell proliferation. The appearance of some inactive cross-reacting material in quiescent cells is probably due to post-translational alteration of the pre-synthesized molecules. Compared with other glycolytic enzymes, such as glucose phosphate isomerase and pyruvate kinase, phosphofructokinase seems to be the (or one of the) preferential target of glycolytic induction in proliferating cells.

Effects of Heparin Oligosaccharides with High Affinity for Antithrombin III in Experimental Venous Thrombosis

1982 ◽  
Vol 47 (03) ◽  
pp. 244-248 ◽  
Author(s):  
D P Thomas ◽  
Rosemary E Merton ◽  
T W Barrowcliffe ◽  
L Thunberg ◽  
U Lindahl
Keyword(s):  
Antithrombin Iii ◽  
Specific Activity ◽  
High Specific Activity ◽  
Factor Xa ◽  
Blood Levels ◽  
Thrombin Time ◽  
High Affinity ◽  
Very High

SummaryThe in vitro and in vivo characteristics of two oligosaccharide heparin fragments have been compared to those of unfractionated mucosal heparin. A decasaccharide fragment had essentially no activity by APTT or calcium thrombin time assays in vitro, but possessed very high specific activity by anti-Factor Xa assays. When injected into rabbits at doses of up to 80 ¼g/kg, this fragment was relatively ineffective in impairing stasis thrombosis despite producing high blood levels by anti-Xa assays. A 16-18 monosaccharide fragment had even higher specific activity (almost 2000 iu/mg) by chromogenic substrate anti-Xa assay, with minimal activity by APTT. When injected in vivo, this fragment gave low blood levels by APTT, very high anti-Xa levels, and was more effective in preventing thrombosis than the decasaccharide fragment. However, in comparison with unfractionated heparin, the 16-18 monosaccharide fragment was only partially effective in preventing thrombosis, despite producing much higher blood levels by anti-Xa assays.It is concluded that the high-affinity binding of a heparin fragment to antithrombin III does not by itself impair venous thrombogenesis, and that the anti-Factor Xa activity of heparin is only a partial expression of its therapeutic potential.

Autoprothrombin C Activity from Prothrombin with Snake Venom or Trypsin

1962 ◽  
Vol 08 (03) ◽  
pp. 425-433 ◽  
Author(s):  
Ewa Marciniak ◽  
Edmond R Cole ◽  
Walter H Seegers
Keyword(s):  
Snake Venom ◽  
Thrombolytic Agent ◽  
Sodium Citrate ◽  
Specific Activity ◽  
High Specific Activity ◽  
Citrate Solution ◽  
Clotting Factors ◽  
Activation Products ◽  
Russell’S Viper ◽  
Autoprothrombin C

SummarySuitable conditions were found for the generation of autoprothrombin C from purified prothrombin with the use of Russell’s viper venom or trypsin. DEAE chromatographed prothrombin is structurally altered and has never been found to yield autoprothrombin C and also did not yield it when Russell’s viper venom or trypsin were used. Autoprothrombin C is derived from prothrombin with tissue extract thromboplastin, but not in large amounts with the intrinsic clotting factors. With the latter thrombin and autoprothrombin III are the chief activation products. Autoprothrombin III concentrates were prepared from serum and upon activation with 25% sodium citrate solution or with Russell’s viper venom large amounts of autoprothrombin C were obtained, and this was of high specific activity. Theoretically trypsin is not a thrombolytic agent, but on the contrary should lead to intravascular clotting.

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