Identification of progesterone receptor membrane component-1 as an interaction partner and possible regulator of fatty acid 2-hydroxylase.

Abstract

The fatty acid 2-hydroxylase (FA2H) is essential for synthesis of 2-hydroxylated fatty acids in myelinating and other cells, and deficiency of this enzyme causes a complicated form of hereditary spastic paraplegia also known as fatty acid hydroxylase-associated neurodegeneration. Despite its important role in sphingolipid metabolism, regulation of FA2H and its interaction with other proteins involved in the same or other metabolic pathways is poorly understood. To identify potential interaction partners of the enzyme, quantitative mass spectrometry using stable isotope labeling of cells was combined with formaldehyde cross-linking and proximity biotinylation, respectively. Besides other enzymes involved in sphingolipid synthesis and intermembrane transfer of ceramide, and putative redox partners of FA2H, progesterone receptor membrane component-1 (PGRMC1) and PGRMC2 were identified as putative interaction partners. These two related heme-binding proteins are known to regulate several cytochrome P450 enzymes. Bimolecular fluorescence complementation experiments confirmed the interaction of FA2H with PGRMC1. Moreover, the PGRMC1 inhibitor AG-205 significantly reduced synthesis of hydroxylated ceramide and glucosylceramide in FA2H-expressing cells. This suggests that PGRMC1 may regulate FA2H activity, possibly through its heme chaperone activity.

• Authors: Robert Hardt, Dominic Winter, Volkmar Gieselmann, Matthias Eckhardt
• Journal: The Biochemical journal (Biochem J) Vol. 475 Issue 5 Pg. 853-871 (03 06 2018) ISSN: 1470-8728 [Electronic] England
• PMID: 29438993 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Chemical References

• Membrane Proteins
• PGRMC1 protein, human
• Receptors, Progesterone
• Heme
• Mixed Function Oxygenases
• fatty acid alpha-hydroxylase

Topics

• Animals
• CHO Cells
• Cells, Cultured
• Cricetinae
• Cricetulus
• Enzyme Activation
• HEK293 Cells
• Heme (metabolism)
• Humans
• Lipid Metabolism
• Membrane Proteins (metabolism, physiology)
• Mixed Function Oxygenases (metabolism)
• Protein Binding
• Receptors, Progesterone (metabolism, physiology)