How to measure LRAT (lecithin-retinol acyltransferase) activity in cell lysate?

08 August 2016 1 6K Report

Interested in measuring converted all-trans retinyl esters in RPE but without using radioactivity.

Trying to find another way to show that the increase of lipid droplets that I see in RPE cells are full of retinyl esters that have been produced via LRAT activity.

Martijn R. Molenaar

If you have a HPLC, you can use a non-endogeneous phospholipid (for example PC(7:0/7:0)) and measure RE-formation with a C7 fatty acid after addition of retinol. Have a look in the reference below. I think you should be able to measure it by TLC as well.

Nat Chem Biol. 2015 Jan;11(1):26-32. doi: 10.1038/nchembio.1687. Epub 2014 Nov 10. LRAT-specific domain facilitates vitamin A metabolism by domain swapping in HRASLS3. Golczak M, Sears AE, Kiser PD, Palczewski K.

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