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H6380

Hexokinase from Saccharomyces cerevisiae

Name: Hexokinase from <I>Saccharomyces cerevisiae</I>
Brand: SIGMA

lyophilized powder, ≥350 units/mg protein, Protein ≥10 % by biuret

Synonym(s):

ATP:D-Hexose-6-phosphotransferase, Hexokinase from yeast

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About This Item

CAS Number:

9001-51-8

UNSPSC Code:

12352204

eCl@ss:

32160410

EC Number:

232-611-5

NACRES:

NA.54

MDL number:

MFCD00131321

EC Number:

2.7.1.1 (BRENDA, IUBMB)

Specific activity:

≥350 units/mg protein

Biological source:

Saccharomyces cerevisiae

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Properties

biological source

Saccharomyces cerevisiae

Quality Segment

200

form

lyophilized powder

specific activity

≥350 units/mg protein

mol wt

dimer 110 kDa

composition

Protein, ≥10% biuret

manufacturer/tradename

Sigma-Aldrich

technique(s)

activity assay: suitable

color

white

pH

UniProt accession no.

P04806, P04807

application(s)

life science and biopharma

foreign activity

L-Glutamic dehydrogenase ≤0.05%, Creatine phosphokinase, myokinase, 6-phosphogluconic dehydrogenase and alcohol dehydrogenase ≤0.001%, Glucose-6-phosphate dehydrogenase ≤0.005%, Glutathione reductase ≤0.005%, Phosphoglucose isomerase ≤0.002%

storage temp.

2-8°C

Gene Information

bakers yeast ... HXK1(850614), HXK2(852639)

Description

General description

Research area: Cell signalling. Hexokinase (HK) from yeast is a 486 amino-acid containing enzyme with a α/β fold. It comprises a large and small domain separated by an active site cleft. Conserved glycine residues are essential for ATP and glucose binding. The two isoforms are HKI and HKII encoded by HXK1 and HXK2 gene, respectively.

Application

Hexokinase from Saccharomyces cerevisiae has been used:

to investigate its amyloid fibril forming tendency using biophysical methods
in tracking ATP depletion in peroxisomes and reticulocyte lysates in import assays
in the synthesis of Mg-ADP-actin from Mg-ATP actin monomers

Biochem/physiol Actions

Hexokinase II is a crucial enzyme in glucose metabolism. Deletion of gene in S. cerevisiae in the presence of high glucose displays oxidative growth and results in the production of high biomass as well as shortened diauxic shift. Hexokinase II is also involved in glucose repression.

Catalyzes the phosphorylation of D-hexose sugars at the C6 position utilizing ATP as a phosphate source.

The rate of phosphorylation varies with different hexoses (pH 7.5, 30 °C).
D-fructose K_M: 0.33 mM
D-glucose K_M: 0.12 mM
D-mannose K_M: 0.05 mM

Yeast hexokinase exists as two similar isoforms, PI and PII (A and B), with isoelectric points of 5.25 and 4, respectively.

Molecular Weight:~54 kDa (monomer)
~110 kDa (dimer)
Optimal pH: 7.5 to 9.0
Extinction Coefficient: E^1% = 8.85 (PI) and 9.47 (PII) at 280 nm

Activators: Hexokinase requires Mg²⁺ ions (K_M = 2.6 mM) for activity. Hexokinase is activated by catecholamines and related compounds.

Inhibitors: sorbose-1-phosphate, polyphosphates, 6-deoxy-6-fluoroglucose, 2-C-hydroxy-methylglucose, xylose, lyxose, and thiol reactive compounds (Hg²⁺ and 4-chloromercuribenzoate)

Physical form

Lyophilized powder containing phosphate/citrate pH approx. 7.0

Other Notes

One unit will phosphorylate 1.0 μmole of D-glucose per min at pH 7.6 at 25 °C, unless otherwise indicated below.

Predominantly the PII isoform, produced from an overexpressing yeast strain.

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Storage Class

11 - Combustible Solids

wgk

WGK 3

flash_point_f

Not applicable

flash_point_c