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Sigma-Aldrich

Hexokinase from Saccharomyces cerevisiae

Type F-300, lyophilized powder, ≥130 units/mg protein (biuret)

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Synonym(s):
Hexokinase D, Hexokinasetype IV, ATP:D-Hexose-6-phosphotransferase, Hexokinase from yeast
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
eCl@ss:
32160410
NACRES:
NA.54

biological source

Saccharomyces cerevisiae

type

Type F-300

form

lyophilized powder

specific activity

≥130 units/mg protein (biuret)

mol wt

110 kDa

purified by

crystallization

storage condition

(Keep container tightly closed in a dry and well-ventilated place)

color

white

anion traces

sulfate (SO42-): free

UniProt accession no.

foreign activity

ATPase, myokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconic dehydrogenase and phosphoglucose isomerase ≤0.01%

storage temp.

−20°C

Gene Information

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

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General description

Research area: Cell Signaling

The hexokinase 1 (HXK1) gene is mapped to the Saccharomyces cerevisiae chromosome VI. HXK1 is ubiquitously expressed in mammalian tissues and is abundant in the brain, erythrocytes, lymphocytes, and fibroblasts.

Application

Hexokinase has been used:
  • for digesting glucose for measuring glucose content in the root and stem samples of Quercus velutina
  • to treat ADP (adenosine diphosphate) solution along with D-glucose to remove the contaminating ATP (adenosine triphosphate)
  • in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (pH 7.5) for ADP-[13CU]glucose synthesis

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 catalyzes the rate-limiting first step of glycolysis, which involves the phosphorylation of glucose.

Quality

A mixture of isoenzymes.

Unit Definition

One unit will phosphorylate 1.0 μmole of D-glucose per min at pH 7.6 at 25 °C.

Physical form

Lyophilized powder containing approx. 15% sodium citrate

Reconstitution

Reconstitute with water or citrate buffer, pH 7.0. Studies have shown excellent stability during repeated freezing and thawing over a period of at least 30 days.

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

Regulatory Information

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Erin Wiley et al.
Tree physiology, 33(11), 1216-1228 (2013-11-26)
There is no consensus about how stresses such as low water availability and temperature limit tree growth. Sink limitation to growth and survival is often inferred if a given stress does not cause non-structural carbohydrate (NSC) concentrations or levels to
Daiki Kubota et al.
Ophthalmic genetics, 41(6), 629-638 (2020-08-21)
The hexokinase 1 (HK1) gene encodes one of the four human hexokinases that play essential roles in glucose metabolism. Recently, several cases of E847K mutation in the HK1 gene were reported to cause inherited retinal dystrophy. The purpose of this
Manfred Schmid et al.
Molecular cell, 21(3), 379-391 (2006-02-04)
Our previous work identified the inner basket of the NPC as a physical activation/protection station for force-tethered, epigenetically silenced genes. Here we show that a specific nucleopore-to-gene-promoter interaction (Nup-PI) is an early physiological event of gene activation. Nup-PI was discovered
RHAU helicase stabilizes G4 in its nucleotide-free state and destabilizes G4 upon ATP hydrolysis
You H, et al.
Nucleic Acids Research, 45(1), 206-214 (2016)
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

Protocols

Measure hexokinase activity using a continuous spectrophotometric rate-determination assay at 340 nm, catalyzing D-hexose sugar phosphorylation using ATP.

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