Chapter title |
Measuring Activity of Cholesterol Synthesis Enzymes Using Gas Chromatography/Mass Spectrometry
|
---|---|
Chapter number | 16 |
Book title |
Cholesterol Homeostasis
|
Published in |
Methods in molecular biology, February 2017
|
DOI | 10.1007/978-1-4939-6875-6_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6873-2, 978-1-4939-6875-6
|
Authors |
Anika V. Prabhu B.Sc., Winnie Luu Ph.D., Andrew J. Brown Ph.D., Prabhu, Anika V., Luu, Winnie, Brown, Andrew J., Anika V. Prabhu, Winnie Luu, Andrew J. Brown |
Editors |
Ingrid C. Gelissen, Andrew J. Brown |
Abstract |
The development of gas chromatography/mass spectrometry (GC/MS) technology has improved the ease and efficiency with which sterols in biological samples can be analyzed. Its advantages include that it needs only a small amount of sample, a short analysis time, and has enhanced specificity over traditional methods. Furthermore, a major benefit is its nonselective properties, which means that a complete scan of the sample will display the relative abundance of every sterol in the sample. This property has made it possible to define the abnormal, but distinctive, sterol profiles in a number of inborn errors of cholesterol synthesis. Here, we describe a semiquantitative method to determine relative activity of cholesterol synthesis enzymes. As an example, we measure the relative abundance of the substrate and product sterols of a cholesterol synthetic enzyme, 24-dehydrocholesterol reductase (DHCR24), which is defective in the hereditary developmental disease desmosterolosis. |
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