Active site mutagenesis of the putative Diels-Alderase macrophomate synthase.
Although the macrophomate synthase active site is rich in potential functional groups, site-directed mutagenesis shows that only three residues are absolutely required for catalysis of oxaloacetate decarboxylation and trapping of the resulting enolate with a 2-pyrone; the other residues that line the binding pocket are surprisingly tolerant to substitution.
Serafimov
Jörg M
JM
Laboratory of Organic Chemistry, ETH Zurich, CH-8093 Zurich, Switzerland.
Lehmann
Hans Christian
HC
Oikawa
Hideaki
H
Hilvert
Donald
D
D001665:Binding Sites; D002384:Catalysis; D008274:Magnesium / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016296:Mutagenesis / Q000235:genetics*; D010071:Oxaloacetates / Q000737:chemistry
D009097:Multienzyme Complexes; D010071:Oxaloacetates; C408144:macrophomate synthase; D008274:Magnesium