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문서

H6380

Sigma-Aldrich

Hexokinase from Saccharomyces cerevisiae

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

동의어(들):

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

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About This Item

CAS Number:
효소 위원회 번호:
EC Number:
MDL number:
UNSPSC 코드:
12352204
eCl@ss:
32160410
NACRES:
NA.54

생물학적 소스

Saccharomyces cerevisiae

Quality Level

형태

lyophilized powder

특이 활성도

≥350 units/mg protein

분자량

dimer 110 kDa

구성

Protein, ≥10% biuret

제조업체/상표

Sigma-Aldrich

기술

activity assay: suitable

색상

white

pH

6

UniProt 수납 번호

응용 분야

life science and biopharma

외래 활성

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%

저장 온도

2-8°C

유전자 정보

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

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일반 설명

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.

애플리케이션

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

생화학적/생리학적 작용

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. Hexokinase II is also involved in glucose repression.

단위 정의

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

물리적 형태

Lyophilized powder containing phosphate/citrate pH approx. 7.0

기타 정보

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

개인 보호 장비

Eyeshields, Gloves, type N95 (US)


시험 성적서(COA)

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문서 라이브러리 방문

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)
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)
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)
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

프로토콜

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

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