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H6380

Sigma-Aldrich

Hexokinase from Saccharomyces cerevisiae

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:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
eCl@ss:
32160410
NACRES:
NA.54

Pricing and availability is not currently available.

biological source

Saccharomyces cerevisiae

Quality Level

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

6

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This Item
D1238496738SML3593
Quality Level

100

Quality Level

200

Quality Level

100

Quality Level

100

storage temp.

2-8°C

storage temp.

-

storage temp.

-

storage temp.

2-8°C

form

powder

form

powder

form

crystals

form

powder

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.[1] The two isoforms are HKI and HKII encoded by HXK1 and HXK2 gene, respectively.[2]

Application

Hexokinase from Saccharomyces cerevisiae has been used:
  • to investigate its amyloid fibril forming tendency using biophysical methods[3]
  • in tracking ATP depletion in peroxisomes and reticulocyte lysates in import assays[4]
  • in the synthesis of Mg-ADP-actin from Mg-ATP actin monomers[5]

Biochem/physiol Actions

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 KM: 0.33 mM
D-glucose KM: 0.12 mM
D-mannose KM: 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: E1% = 8.85 (PI) and 9.47 (PII) at 280 nm

Activators: Hexokinase requires Mg2+ ions (KM = 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)
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.[2] Hexokinase II is also involved in glucose repression.[2]

Unit Definition

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

Physical form

Lyophilized powder containing phosphate/citrate pH approx. 7.0

Other Notes

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

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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    The high resolution crystal structure of yeast hexokinase PII with the correct primary sequence provides new insights into its mechanism of action
    Kuser PR, et al.
    Test, 275(27), 20814-20821 (2000)
    Large proteins have a great tendency to aggregate but a low propensity to form amyloid fibrils
    Ramshini H, et al.
    Testing, 6(1), e16075-e16075 (2011)
    The energetics of Pex5p-mediated peroxisomal protein import
    Oliveira M, et al.
    Test, 278(41), 39483-39488 (2003)
    Mechanism of actin polymerization revealed by cryo-EM structures of actin filaments with three different bound nucleotides
    Chou SZ and Pollard TD
    Proceedings of the National Academy of Sciences of the USA, 116(10), 4265-4274 (2019)
    J A Diderich et al.
    Applied and environmental microbiology, 67(4), 1587-1593 (2001-04-03)
    Hexokinase II is an enzyme central to glucose metabolism and glucose repression in the yeast Saccharomyces cerevisiae. Deletion of HXK2, the gene which encodes hexokinase II, dramatically changed the physiology of S. cerevisiae. The hxk2-null mutant strain displayed fully oxidative

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