H6380

Hexokinase from Saccharomyces cerevisiae

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

• Sinonimo/i: ATP:D-Hexose-6-phosphotransferase, Hexokinase from yeast
• Numero CAS: 9001-51-8
• Classificazione EC (Enzyme Commission): 2.7.1.1 (BRENDA, IUBMB)
• Numero CE: 232-611-5
• Numero MDL: MFCD00131321
• Codice UNSPSC: 12352204
• eCl@ss: 32160410
• NACRES: NA.54

Proprietà

• Origine biologica: Saccharomyces cerevisiae
• Livello qualitativo: 200
• Forma fisica: lyophilized powder
• Attività specifica: ≥350 units/mg protein
• PM: dimer 110 kDa
• Composizione: Protein, ≥10% biuret
• Produttore/marchio commerciale: Sigma-Aldrich
• tecniche: activity assay: suitable
• Colore: white
• pH: 6
• N° accesso UniProt: P04806; P04807
• applicazioni: life science and biopharma
• Attività estranea: 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%
• Temperatura di conservazione: 2-8°C
• Informazioni sul gene: bakers yeast ... HXK1(850614) , HXK2(852639)

Descrizione

Descrizione generale

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.

Applicazioni

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

Azioni biochim/fisiol

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 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.

Definizione di unità

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

Stato fisico

Lyophilized powder containing phosphate/citrate pH approx. 7.0

Altre note

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

Informazioni sulla sicurezza

• Codice della classe di stoccaggio: 11 - Combustible Solids
• Classe di pericolosità dell'acqua (WGK): WGK 3
• Punto d’infiammabilità (°F): Not applicable
• Punto d’infiammabilità (°C): Not applicable
• Dispositivi di protezione individuale: Eyeshields, Gloves, type N95 (US)