Site-directed mutagenesis of a fatty acid elongase ELO-like condensing enzyme

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• 作者：Selene Hern ; ez-Buquer ; Brenda J. Blacklock
• 关键词：VLCFA ; very long chain fatty acids ; CoA ; coenzyme A ; KCS ; 3-ketoacylCoA synthase ; GC/MS ; gas chromatography/mass spectrometry ; ER ; endoplasmic reticulum
• 刊名：FEBS Letters
• 出版年：29 November, 2013
• 年：2013
• 卷：587
• 期：23
• 页码：3837-3842
• 全文大小：1903 K

文摘

The condensation step of fatty acid elongation is the addition of a C₂ unit from malonyl-CoA to an acyl primer catalyzed by one of two families of enzymes, the 3-ketoacyl-CoA synthases and the ELO-like condensing enzymes. 3-Ketoacyl-CoA synthases use a Claisen-like reaction mechanism while the mechanism of the ELO-catalyzed condensation reaction is unknown. We have used site-directed mutagenesis of Dictyostelium discoideum EloA to identify residues important to catalytic activity and/or structure. Mutation of highly conserved polar residues to alanine resulted in an inactive enzyme strongly suggesting that these residues play a role in the condensation reaction.