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Lipid specificity and location of the sterol carrier protein-2 fatty acid-binding site: A fluorescence displacement and energy transfer study

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Lipids

Abstract

Although it was recently recognized that sterol carrier protein-2 (SCP-2) interacts with fatty acids, little is known regarding the specificity of SCP-2 for long-chain fatty acids or branched-chain fatty-acid-like molecules. Likewise the location of the fatty-acid binding site within SCP-2 is unresolved. A fluorescent cis-parinaric acid displacement assay was used to show that SCP-2 optimally interacted with 14–22 carbon chain lipidic molecules: polyunsaturated fatty acids > monounsaturated, saturated > branched-chain isoprenoids > branched-chain phytol-derived fatty acids. In contrast, the other major fatty-acid binding protein in liver, fatty-acid binding protein (L-FABP), displayed a much narrower carbon chain preference in general: polyunsaturated fatty acids > branched-chain phytol-derived fatty acids > 14- and 16-carbon saturated > branched-chain isoprenoids. However, both SCP-2 and l-FABP displayed a very similar unsaturated fatty-acid specificity profile. The presence and location of the SCP-2 lipid binding site were investigated by fluorescence energy transfer. The distance between the SCP-2 Trp50 and bound cis-parinaric acid was determined to be 40 Å. Thus, the SCP-2 fatty-acid binding site appeared to be located on the opposite side of the SCP-2 Trp50. These findings not only contribute to our understanding of the SCP-2 ligand binding site but also provide evidence suggesting a potential role for SCP-2 and/or L-FABP in metabolism of branched-chain fatty acids and isoprenoids.

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Abbreviations

SCP-2:

sterol carrier protein-2

L-FABP:

liver fatty-acid binding protein

NMR:

nuclear magnetic resonance

cis-parinaric acid:

9Z,11E,13E,15Z-octadecatetraenoic acid

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Authors and Affiliations

  1. Department of Physiology and Pharmacology, Texas A&M University, TVMC, 77843-4466, College Station, Texas

    Andrey Frolov, Kimberly Miller, Tae-Hyeon Cho & Friedhelm Schroeder

  2. Cardiovascular Department, DuPont Merck Pharmaceutical Company, Experimental Station 400-3231, 19898-0400, Wilmington, Delaware

    Jeffrey T. Billheimer

Authors
  1. Andrey Frolov
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  2. Kimberly Miller
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  3. Jeffrey T. Billheimer
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  4. Tae-Hyeon Cho
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  5. Friedhelm Schroeder
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Correspondence to Friedhelm Schroeder.

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This work was supported in part by grants from the United States Public Health Service National Institutes of Health (DK41402 and GM31651).

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Frolov, A., Miller, K., Billheimer, J.T. et al. Lipid specificity and location of the sterol carrier protein-2 fatty acid-binding site: A fluorescence displacement and energy transfer study. Lipids 32, 1201–1209 (1997). https://doi.org/10.1007/s11745-997-0154-5

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  • DOI: https://doi.org/10.1007/s11745-997-0154-5

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