The inhibition of glucokinase and glycerokinase from <i>Bacillus stearothermophilus</i> by the triazine dye Procion Blue MX-3G

Goward, Christopher R.; Scawen, Michael D.; Atkinson, Tony

doi:10.1042/bj2460083

Cited by 13 publications

(6 citation statements)

References 21 publications

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“…stearothermophilus used here is much smaller in size (MW about 30,000) compared with mammalian GK (MW of approximately 50,000). It is also smaller than Zymomonas mobilis GK (Goward et al, 1987) which has an estimated MW of 38,000 (Barnell et al, 1990). cDNA sequence is not available for B. stearothermophilus GK but available for Z .…”

Section: Discussionmentioning

confidence: 99%

Interaction of facilitative glucose transporter with glucokinase and its modulation by ADP and glucose‐6‐phosphate

Lachaal¹,

Jung

1993

Journal Cellular Physiology

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Bacterial glucokinase (GK) binds to purified, human erythrocyte glucose transporter (GT) reconstituted in vesicles. The binding is largely abolished if GT is predigested with trypsin, indicating that GK binds to the cytoplasmic domain of GT. The binding is a saturable function of GK concentration showing two distinct affinities with apparent KD of 0.33 and 5.1 microM. The binding is stimulated by an increasing concentration of ADP with the 50% maximal effect at 5 mM. Glucose-6-phosphate (G6P) also stimulates the binding with a distinct optimum at 25 mM. The binding is stimulated only slightly by ATP. D-glucose has no affect on the binding. KCl enhances the binding with the maximal effect at physiological intracellular concentrations. The binding is sensitive to changes in pH with an optimum at pH 4. The binding causes no detectable functional change in GT. However, the enzymatic activity of GK measured at nanomolar concentrations of GK is significantly greater in the presence of GT vesicles than in its absence or in the presence of protein-free vesicles, indicating that GK interacts with GT at this low concentration range with an apparent KD of 10 mM. Although its physiological significance is not known, the GK-GT interaction in vitro described here suggests that these two proteins may also interact in the cell and regulate carbohydrate metabolism.

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Section: Discussionmentioning

confidence: 99%

Interaction of facilitative glucose transporter with glucokinase and its modulation by ADP and glucose‐6‐phosphate

Lachaal¹,

Jung

1993

Journal Cellular Physiology

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“…These enzymes are already in use as biocatalysts in industrial processes (Jaenicke et al 1996;Sthal 1993). A glucokinase from the thermophilic organism Bacillus stearothermophilus has been characterized and is known to display long term stability (Goward et al 1987;Ishikawa et al 1987;Tomita et al 1990). Hence we evaluated the use of this glucokinase (BSGK) in the absence of ATP as a non-consuming glucose sensor.…”

Section: Thermostable Glucose Dehydrogenase From Thermoplasma Acidophmentioning

confidence: 99%

Thermostable Proteins as Probe for the Design of Advanced Fluorescence Biosensors

Champdoré

Staiano

Aurilia

et al. 2006

Rev Environ Sci Biotechnol

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In this review we explore the advantages deriving from the use of either enzymes or sugar binding proteins isolated from thermophilic organisms to develop stable fluorescence biosensors. We report on a novel approach to address the consumption of the analyte by enzyme-based biosensors, namely the utilization of apo-enzymes as non-active forms of proteins which are still able to bind the ligand but cannot transform it into product. We also report recent studies in which the fluorescence labeling of a naturally thermostable binding protein allows a quantitative determination of glucose. Abbreviations IAANS 2-(4¢-(iodoacetamido)anilino)naphthalene-6-sulfonic acid (IAANS) ANS 1-(anilino)-naphtalene-8-sulfonate FRET fluorescence resonance energy transfer LED light emitting diode GD glucose dehydrogenase BSGK glucokinase from Bacillus stearothermophilus Ph-SBP sugar-binding protein from Pyrococcus horikoshii M. de Champdoré AE M. Staiano AE V. Aurilia AE

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“…A glucokinase from the thermophilic organism Bacillus stearothermophilus has been characterized (56,57) and is known to display long-term stability (52). Hence we evaluated the use of this glucokinase (BSGK) in the absence of ATP as a glucose-nonconsuming glucose sensor.…”

Section: Structure and Mechanism Of Glucose Binding Of Hexokinasementioning

confidence: 99%

A Novel Fluorescence Competitive Assay for Glucose Determinations by Using a Thermostable Glucokinase from the Thermophilic Microorganism Bacillus stearothermophilus

D’Auria

DiCesare

Staiano

et al. 2002

Analytical Biochemistry

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We describe the use of a thermostable glucokinase in a novel competitive fluorescence assay for glucose. Glucokinase from Bacillus stearothermophilus (BSGK) was found to retain enzymatic activity in solution for over 20 days. The single cysteine residue in BSGK, which is near the active site, was labeled with a fluorescent probe, 2-(4-iodoacetamidoanilino)naphthalene-6-sulfonic acid. The ANS-labeled BSGK displayed a modest 25% decrease in the emission intensity upon binding glucose but no change in lifetime. To obtain a larger spectral change we developed a competitive assay for glucose using the intrinsic tryptophan fluorescence from BSGK and a resonance energy transfer (RET) acceptor-labeled sugar. The sugar-labeled acceptor quenched the BSGK tryptophan emission, and the quenching was reversed upon addition of glucose. The use of RET as the sensing mechanism can be easily extended to longer wavelengths for a more practical glucose sensor.

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The inhibition of glucokinase and glycerokinase from Bacillus stearothermophilus by the triazine dye Procion Blue MX-3G

Cited by 13 publications

References 21 publications

Interaction of facilitative glucose transporter with glucokinase and its modulation by ADP and glucose‐6‐phosphate

Interaction of facilitative glucose transporter with glucokinase and its modulation by ADP and glucose‐6‐phosphate

Thermostable Proteins as Probe for the Design of Advanced Fluorescence Biosensors

A Novel Fluorescence Competitive Assay for Glucose Determinations by Using a Thermostable Glucokinase from the Thermophilic Microorganism Bacillus stearothermophilus

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