Cloning, Purification, and Characterization of the Catalytic C-Terminal Domain of the Human 3-Hydroxy-3-methyl glutaryl-CoA Reductase: An Effective, Fast, and Easy Method for Testing Hypocholesterolemic Compounds.
Amino Acid Sequence
/ genetics
Catalytic Domain
Chromatography, Affinity
Cloning, Molecular
Drug Evaluation, Preclinical
/ methods
Enzyme Assays
/ methods
Escherichia coli
/ genetics
Gene Expression
Humans
Hydroxymethylglutaryl CoA Reductases
/ chemistry
Hydroxymethylglutaryl-CoA Reductase Inhibitors
/ chemistry
Recombinant Proteins
/ chemistry
Tandem Mass Spectrometry
Affinity chromatography
Bacterial expression
Enzymatic activity
HMGR
MALDI MS and MS/MS
Screening of statin-like molecules
Journal
Molecular biotechnology
ISSN: 1559-0305
Titre abrégé: Mol Biotechnol
Pays: Switzerland
ID NLM: 9423533
Informations de publication
Date de publication:
Feb 2020
Feb 2020
Historique:
pubmed:
24
11
2019
medline:
10
6
2020
entrez:
24
11
2019
Statut:
ppublish
Résumé
3-hydroxy-3-methyl glutaryl-CoA reductase, also known as HMGR, plays a crucial role in regulating cholesterol biosynthesis and represents the main pharmacological target of statins. In mammals, this enzyme localizes to the endoplasmic reticulum membrane. HMGR includes different regions, an integral N-terminal domain connected by a linker-region to a cytosolic C-terminal domain, the latter being responsible for enzymatic activity. The aim of this work was to design a simple strategy for cloning, expression, and purification of the catalytic C-terminal domain of the human HMGR (cf-HMGR), in order to spectrophotometrically test its enzymatic activity. The recombinant cf-HMGR protein was heterologously expressed in Escherichia coli, purified by Ni
Identifiants
pubmed: 31758489
doi: 10.1007/s12033-019-00230-1
pii: 10.1007/s12033-019-00230-1
doi:
Substances chimiques
Hydroxymethylglutaryl-CoA Reductase Inhibitors
0
Recombinant Proteins
0
Hydroxymethylglutaryl CoA Reductases
EC 1.1.1.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
119-131Subventions
Organisme : Associazione Italiana per la Ricerca sul Cancro
ID : 15404/2014
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