H4502

Hexokinase from Saccharomyces cerevisiae

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

• Synonym(s): Hexokinase D, Hexokinasetype IV, ATP:D-Hexose-6-phosphotransferase, Hexokinase from yeast
• CAS Number: 9001-51-8
• Enzyme Commission number: 2.7.1.1 (BRENDA, IUBMB)
• EC Number: 232-611-5
• MDL number: MFCD00131321
• UNSPSC Code: 12352204
• eCl@ss: 32160410
• NACRES: NA.54

Properties

• biological source: Saccharomyces cerevisiae
• Quality Level: 200
• 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 (SO₄^2-): free
• UniProt accession no.: P04806; P04807
• 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)

Description

General description

Research area: Cell Signaling

The hexokinase 1 (HXK1) gene is mapped to the Saccharomyces cerevisiae chromosome VI.[1] HXK1 is ubiquitously expressed in mammalian tissues and is abundant in the brain, erythrocytes, lymphocytes, and fibroblasts.[2]

Application

Hexokinase has been used:

• for digesting glucose for measuring glucose content in the root and stem samples of Quercus velutina[3]
• to treat ADP (adenosine diphosphate) solution along with D-glucose to remove the contaminating ATP (adenosine triphosphate)[4]
• in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (pH 7.5) for ADP-[13CU]glucose synthesis[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 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)

Hexokinase catalyzes the rate-limiting first step of glycolysis, which involves the phosphorylation of glucose.[6]

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.

Safety Information

• 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