Differential effects of cholesterol and its immediate biosynthetic precursors on membrane organization. Biochemistry 47: 5668–5677

ABSTRACT: Cholesterol is the most representative sterol present in vertebrate membranes and is the end product of the long and multistep sterol biosynthetic pathway. 7-Dehydrocholesterol (7-DHC) and desmosterol are the immediate biosynthetic precursors of cholesterol in the Kandutsch-Russell and Bloch pathway. In this article, we have monitored the effect of cholesterol and its two immediate biosynthetic precursors on biophysical and dynamic properties of fluid and gel phase membranes. Toward this goal, we have used fluorescent membrane probes, DPH and TMA-DPH, and the hydrophobic probe, pyrene. Our results using these probes show that although both 7-DHC and desmosterol differ with cholesterol in one double bond, they exhibit differential effects on membrane organization and dynamics. Importantly, we show that the effect of cholesterol and desmosterol on membrane organization and dynamics is similar in most cases, while 7-DHC has a considerably different effect. This demonstrates that the position of the double bond in sterols is an important determinant in maintaining membrane order and dynamics. These results assume relevance since the accumulation of cholesterol precursors have been reported to result in severe pathological conditions. Cholesterol is an essential constituent of eukaryotic membranes and plays a crucial role in membrane organiza