PMID: 8457568Mar 30, 1993Paper

Affinity labeling of lecithin retinol acyltransferase

Biochemistry
Y Q ShiR R Rando

Abstract

Lecithin retinol acyltransferase (LRAT) transfers acyl groups regiospecifically from the sn-1 position of lecithins to all-trans-retinol (vitamin A) and similar retinoids. LRAT is essential for the biosynthesis of 11-cis-retinal, the visual pigment chromophore. LRAT is also required for the general dietary mobilization of vitamin A. The enzyme is membrane-bound and has been solubilized and partially, but not completely, purified. It is demonstrated here that all-trans-retinyl alpha-bromoacetate (RBA) is a potent irreversible affinity labeling agent of LRAT. The measured KI = 12.1 microM and the pseudo-first-order rate constant for inhibition is kinh = 8.2 x 10(-4) s-1. The specificity of the inhibition process is further evidenced by the observation that alpha-bromoacetate derivatives of hydrophobic alcohols which are not substrates for LRAT, such as cholesterol and beta-ionol, are not inhibitors of the enzyme. Labeling of the partially purified enzyme with 3H-RBA showed a single radiolabeled band of molecular weight approximately 25,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

References

Mar 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·O L Francone, C J Fielding
Oct 14, 1988·Biochemical and Biophysical Research Communications·P N MacDonald, D E Ong
Apr 13, 1982·Biochemistry·M A Gawinowicz, D S Goodman
Feb 9, 1993·Biochemistry·Y Q ShiR R Rando

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Citations

Jun 8, 2001·Progress in Retinal and Eye Research·J K McBeeD R Pepperberg
Oct 16, 1999·Biochimica Et Biophysica Acta·J L Napoli
Sep 23, 2003·Biochimica Et Biophysica Acta·Deviprasad R Gollapalli, Robert R Rando
Aug 31, 2006·Biochemistry·Linlong XueRobert R Rando
Jun 26, 2004·Proceedings of the National Academy of Sciences of the United States of America·Deviprasad R Gollapalli, Robert R Rando
Feb 12, 2000·Pharmaceutical Development and Technology·Y Asai, S Watanabe
Aug 2, 2005·Experimental Eye Research·Catherine H KaschulaGabriel H Travis
May 18, 2016·Biochemistry·Avery E Sears, Krzysztof Palczewski
Oct 12, 2005·The Journal of Biological Chemistry·Marcin GolczakKrzysztof Palczewski
Jan 28, 1999·The Journal of Biological Chemistry·A RuizD Bok
Dec 17, 2003·Biochemistry·Deviprasad R Gollapalli, Robert R Rando
Nov 17, 2001·Chemical Reviews·R R Rando
Apr 24, 2002·Biochemical and Biophysical Research Communications·Robert R Rando
May 16, 2002·Biochemistry·Wan Jin JahngRobert R Rando
May 14, 2003·Biochemistry·Deviprasad R Gollapalli, Robert R Rando

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