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Cloning and functional expression of the cDNA encoding rat lanosterol 14-alpha demethylase
1Syntex Discovery Research, Palo Alto, CA 94303, USA.
Gene
|August 19, 1995
Summary
Researchers isolated the full-length coding sequence for rat Lanosterol 14 alpha-demethylase (LDM), a key enzyme in cholesterol synthesis. This discovery aids in developing new cholesterol-lowering drugs by enabling further study of LDM
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Lanosterol 14 alpha-demethylase (LDM) is a cytochrome P-450 enzyme crucial for cholesterol biosynthesis.
- LDM is a potential target for developing cholesterol-lowering drugs.
Purpose of the Study:
- To isolate and characterize the full-length coding sequence of rat LDM (rLDM).
- To facilitate research into cholesterol regulation and drug discovery.
Main Methods:
- Purification of rLDM from cholestyramine-treated rat livers.
- Tryptic fragmentation, amino acid sequencing, and RT-PCR for gene cloning.
- Expression of rLDM in a baculovirus/insect cell system.
Main Results:
- Isolation of a clone encoding a 486 amino acid polypeptide for rLDM.
- Deduced amino acid sequence shows high identity to yeast LDM and conserved P-450 motifs.
- Expressed rLDM exhibited enzymatic activity inhibited by azalanstat (IC50 < 2 nM).
Conclusions:
- The full-length rLDM coding sequence has been successfully isolated and characterized.
- This provides a valuable tool for studying cholesterol biosynthesis regulation.
- Enables further investigation into LDM as a target for novel cholesterol-lowering therapies.