Site-directed mutagenesis of a fatty acid elongase ELO-like condensing enzyme |
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Authors: | Selene Hernandez-Buquer Brenda J Blacklock |
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Institution: | Department of Chemistry and Chemical Biology, Indiana University – Purdue University Indianapolis, Indianapolis, IN 46202, USA |
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Abstract: | 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. |
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Keywords: | VLCFA very long chain fatty acids CoA coenzyme A KCS 3-ketoacylCoA synthase GC/MS gas chromatography/mass spectrometry ER endoplasmic reticulum |
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