Pig red blood cell hexokinase: Evidence for the presence of hexokinase types II and III, and their purification and characterization

Stocchi, Vilberto; Magnani, Mauro; Novelli, Giuseppe; Dachà, Marina; Fornaini, G

doi:10.1016/0003-9861(83)90303-x

Cited by 24 publications

(14 citation statements)

References 19 publications

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“…The overall loss of activity in the absence of fructose following fractionation was almost 70%, and in the presence of fructose, about 25% (data not shown). The use of fructose to stabilize mammalian hexokinases has been reported (15,27) but, to our knowledge, problems with FK stability have not been reported previously for higher plants. The purification protocol resulted in a 700-fold increase in specific activity.…”

Section: Resultsmentioning

confidence: 85%

Purification and Properties of Fructokinase from Developing Tubers of Potato (Solanum tuberosum L.)

Gardner¹,

Davies²,

Burch³

1992

Plant Physiol.

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Fructokinase has been purified from developing potato (Solanum tuberosum L.) tubers by a combination of hydrophobic interaction, affinity chromatography, and gel filtration. The protein has a native molecular mass of approximately 70 kD but is apparently a dimer. Ion-exchange chromatography and two-dimensional western blots resolved three major fructokinases, designated FK-1, FK-11, and FKIlIl in order of their elution from a Mono-Q column. Fructokinase activity proved labile when proteins were purified in the absence of fructose. Kinetically, FKs I, Il, and IlIl all have broad pH optima with peaks at about pH 8.5. The enzymes have a high specificity for fructose (Km values ranging from 0.041 to 0.128 mM), and can utilize a range of nucleoside triphosphates. Unlike FKs I and 11, FK-III is not inhibited by fructose concentrations in excess of 1 mm. MgADP inhibited activity of the three FKs (between 68 and 75% inhibition at 1.0 mM), whereas fructose 6-P caused inhibition at concentrations of 10 mm. There were no regulatory effects observed with a range of other metabolites. K' (10 mM) activated FK-I by 4-fold and FKs II and IlIl by only about 50%.

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Section: Resultsmentioning

confidence: 85%

Purification and Properties of Fructokinase from Developing Tubers of Potato (Solanum tuberosum L.)

Gardner¹,

Davies²,

Burch³

1992

Plant Physiol.

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“…Unit 4 was used for the control sample. RBCs were resuspended in 0.9% (w/v) NaCl in the presence hexokinase was the only enzyme of the glycolytic pathway to be inactivated [Stocchi et al, 1994[Stocchi et al, , 1995. or absence of ascorbate and iron and were exposed to a 50 Hz (0.2 -0.5 mT) magnetic field for up to 90 min Moreover, we also observed a modification in energy charge, a depletion of GSH content, and increased met-at 37 ЊC.…”

Section: Introductionmentioning

confidence: 94%

“…The evidence for adverse health quencies and strengths are produced by high-voltage effects of 50 and 60 Hz magnetic fields [Wertheimer power lines, household appliances, radio and television and Leeper, 1979;Tomenius, 1986;Savitz et al, 1988] antennas, radars, and industrial equipment. Several and the discovery that sinusoidal extremely-low-frestudies reported in the literature have found evidence quency (ELF) electric and magnetic fields affect RNA for a relationship between the exposure to EMFs and transcription and protein synthesis [Goodman and Henthe increased incidence of certain types of tumor, par-derson, 1986, 1988Goodman et al, 1989Goodman et al, , 1993, ticularly leukemia and brain cancer [Wertheimer and transformation of mytotic into nonmytotic cells [RodeLeeper, 1979[RodeLeeper, , 1982Lester and Moore, 1982;Miham, mann et al, 1989], cellular signal transduction [Eich-1982;Lin et al, 1985;Preston-Martin et al, 1987;wald and Kaiser, 1995] and, moreover, that they may Bastuj-Garin et al, 1990;Loomis and Savitz, 1990;Feychting and Ahlbom, 1993;Wertheimer et al, 1995]. However, to date, the results from these studies have *Correspondence to: Prof. Vilberto Stocchi, Istituto di Chimica Biologica ''Giorgio Fornaini,'' Università degli Studi di Urbino, via Saffi 2, not been entirely confirmed by other authors [Fulton 61029 Urbino (PS), Italy.…”

Section: Introductionmentioning

confidence: 99%

In vitro effects of 50 Hz magnetic fields on oxidatively damaged rabbit red blood cells

Fiorani

Biagiarelli

Vetrano

et al. 1997

Bioelectromagnetics

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“…Stored porcine erythrocytes do not utilize glucose during storage unlike stored human erythrocytes for two reasons; first porcine erythrocytes, unlike other mammalian erythrocytes, contain a hexokinase III, which is responsible for 98% of the total glucose phosphorylating activity. 29 The presence of this hexokinase reduces the erythrocyte cell membranes ability to transport glucose and therefore, porcine erythrocytes have a reduced capacity to metabolize glucose. 30 Second, the starting acidic pH inhibits relevant enzyme systems.…”

Section: Perioperative Medicinementioning

confidence: 99%

Interactions of Cardiopulmonary Bypass and Erythrocyte Transfusion in the Pathogenesis of Pulmonary Dysfunction in Swine

Patel¹,

Lin²,

Jones

et al. 2013

Anesthesiology

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Background: Allogeneic erythrocyte transfusion in cardiac surgical patients is associated with a fourfold increase in pulmonary complications. Our understanding of the processes underlying these observations is poor and there is no experimental model of transfusion-related acute lung injury that shows homology to cardiac surgical patients. Our objective was to develop a novel swine recovery model to determine how two clinical risk factors, allogenic erythrocyte transfusion and cardiopulmonary bypass, interact in the genesis of postcardiac surgery acute lung injury. Methods: Thirty-six pigs were infused with allogeneic 14-or 42-day-old erythrocytes or they underwent cardiopulmonary bypass with or without transfusion of 42-day erythrocyte. Controls received saline. All pigs were recovered and assessed for pulmonary dysfunction, inflammation, and endothelial activation at 24 h. Results: Transfusion of stored allogeneic erythrocytes in pigs compared with sham caused pulmonary dysfunction characterized by reduced lung compliance (mean difference −3.36 [95% CI, −5.31 to −1.42] ml/cm H 2 O), an increase in protein levels in bronchoalveolar lavage fluid, histological lung injury inflammation, and endothelial activation. Transfusion of blood stored for up to 42 days resulted in greater protein levels in bronchoalveolar lavage fluid, macrophage infiltration, platelet activation, and depletion of T-lymphocytes in recipient lungs versus 14-day-old blood. Transfusion interacted with cardiopulmonary bypass to increase lung injury in the absence of platelet activation. Conclusions: In this novel large animal model of allogeneic erythrocyte transfusion, pulmonary dysfunction occurs in the absence of any priming event, is increased when combined with other inflammatory stimuli, and is mediated by monocyte activation and T-lymphocyte depletion. What We Already Know about This Topic• Transfusion of allogeneic erythrocytes increases the risk of pulmonary morbidity postcardiac surgery.• Using a novel in vivo porcine model of pulmonary dysfunction, this study determined whether (1) the transfusion of older erythrocytes would cause greater pulmonary dysfunction compared with younger erythrocytes; and (2) erythrocyte transfusion would interact with cardiopulmonary bypass to increase the severity of pulmonary dysfunction. What This Article Tells Us That Is New• Allogeneic erythrocyte transfusion of older erythrocytes causes pulmonary dysfunction, which is characterized by marked neutrophil/macrophage infiltration. Moreover, transfusion interacted with cardiopulmonary bypass to increase lung injury.

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Pig red blood cell hexokinase: Evidence for the presence of hexokinase types II and III, and their purification and characterization

Cited by 24 publications

References 19 publications

Purification and Properties of Fructokinase from Developing Tubers of Potato (Solanum tuberosum L.)

Purification and Properties of Fructokinase from Developing Tubers of Potato (Solanum tuberosum L.)

In vitro effects of 50 Hz magnetic fields on oxidatively damaged rabbit red blood cells

Interactions of Cardiopulmonary Bypass and Erythrocyte Transfusion in the Pathogenesis of Pulmonary Dysfunction in Swine

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