Requirement for a major soluble protein in the conversion of lanosterol to cholesterol by membrane-bound enzymes

Abstract

The conversion of the 30-carbon atom sterol, lanosterol, to cholesterol by a series of membrane-bound rat liver enzymes requires one major soluble protein called squalene and sterol carrier protein (SCP). This homogenous low-molecular-weight liver protein was previously known to function with membrane-bound enzymes catalyzing cholesterol synthesis from 27-carbon atom precursor sterols. To define characteristics of the multienzyme system catalyzing lanosterol metabolism and the role of SCP in this process, a rapid spectroscopic assay was developed, i.e., formation of Δ 5,7-cholestadienol from lanosterol. In addition to SCP, the cofactor requirements for synthesis of cholesterol from lanosterol are NAD, NADPH, and oxygen. Metal ions, reducing agents, heme, or heme-containing proteins are not required. Another homogeneous, low-molecular-weight protein, which accompanies SCP during purification steps, does not support sterol metabolism by membrane-bound enzymes. The broad functions of SCP in cholesterol synthesis and metabolism coupled with its remarkable abundance (~8% of the liver-soluble proteins), ubiquitous occurrence, and recently discovered functions in fatty acid metabolism suggest SCP plays an important regulatory role in lipid metabolism.