title: Catalytic action of fuculose 1-phosphate aldolase (class II) as derived from structure-directed mutagenesis.
abstract: Previous analyses established the structures of unligated L-fuculose 1-phosphate aldolase and of the enzyme ligated with an inhibitor mimicking the substrate dihydroxyacetone phosphate. These data allowed us to suggest a catalytic mechanism. On the basis of this proposal, numerous mutations were now introduced at the active center and tested with respect to their catalytic rates and their product distributions. For several mutants, the structures were determined. The results demonstrate the catalytic importance of some particular residues in defined conformations and in the mobile C-terminal chain end. Moreover, they led to a modification of the proposed mechanism. The effect of some mutations on enantioselectivity and on the ratio of diastereomer formation indicates clearly the binding site of the aldehyde moiety in relation to the other substrate dihydroxyacetone phosphate.
authors.affiliation: Institut für Organische Chemie und Biochemie, Albert-Ludwigs-Universität, D-79104 Freiburg im Breisgau, Germany.
authors.forename: A C
authors.initials: AC
authors.lastname: Joerger
authors.forename: C
authors.initials: C
authors.lastname: Gosse
authors.forename: W D
authors.initials: WD
authors.lastname: Fessner
authors.forename: G E
authors.initials: GE
authors.lastname: Schulz
mesh_terms: D000446:Aldehyde-Lyases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D000447:Aldehydes / Q000737:chemistry; D019943:Amino Acid Substitution / Q000235:genetics; D001426:Bacterial Proteins; D001665:Binding Sites / Q000235:genetics; D002384:Catalysis; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins; D005634:Fructose-Bisphosphate Aldolase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006877:Hydroxamic Acids / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D015032:Zinc / Q000737:chemistry / Q000378:metabolism
chemical_list: D000447:Aldehydes; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D006877:Hydroxamic Acids; D008024:Ligands; D010446:Peptide Fragments; C008886:phosphoglycolohydroxamate; D000446:Aldehyde-Lyases; C103622:FucA protein, bacteria; C491231:fucA protein, E coli; D005634:Fructose-Bisphosphate Aldolase; C036027:L-fuculosephosphate aldolase; D015032:Zinc