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Site‐directed mutagenesis of a fatty acid elongase ELO‐like condensing enzyme
Author(s) -
Hernandez-Buquer Selene,
Blacklock Brenda J.
Publication year - 2013
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2013.10.011
Subject(s) - mutagenesis , enzyme , site directed mutagenesis , chemistry , biochemistry , stereochemistry , directed mutagenesis , primer (cosmetics) , saturated mutagenesis , claisen condensation , catalysis , mutation , organic chemistry , gene , mutant
The condensation step of fatty acid elongation is the addition of a C 2 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.