title: The C-terminal domain of peptide deformylase is disordered and dispensable for activity.
abstract: Upon trypsinolysis, the 18 C-terminal residues of Escherichia coli peptide deformylase were removed but the resulting form exhibited full activity. Moreover, a mutant fms gene encoding the first 145 out of the 168 residues of the enzyme was able to complement a fms(Ts) strain and exhibited full activity. Upon progressive truncation up to residue 139, both activity and stability decreased up to complete inactivation. Mutagenesis of residues of the 138-145 region highlights the importance of Leu-141 and Phe-142. N-Terminal deletions were also carried out. Beyond two residues off, the enzyme showed a dramatic instability. Finally, NMR and thermostability studies of the full-length enzyme and comparison to the 1-147 form strongly suggest that the dispensable residues are disordered in solution.
authors.affiliation: Laboratoire de Biochimie, Unité de Recherche Associée No. 1970 du Centre National de la Recherche Scientifique, Ecole Polytechnique, Palaiseau, France.titi@botrytis.polytechnique.fr
authors.forename: T
authors.initials: T
authors.lastname: Meinnel
authors.forename: C
authors.initials: C
authors.lastname: Lazennec
authors.forename: F
authors.initials: F
authors.lastname: Dardel
authors.forename: J M
authors.initials: JM
authors.lastname: Schmitter
authors.forename: S
authors.initials: S
authors.lastname: Blanquet
mesh_terms: D000581:Amidohydrolases; D000626:Aminopeptidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001665:Binding Sites; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology; D005816:Genetic Complementation Test; D006358:Hot Temperature; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D014357:Trypsin; D015032:Zinc / Q000032:analysis
chemical_list: D010446:Peptide Fragments; D000626:Aminopeptidases; D014357:Trypsin; D000581:Amidohydrolases; C084507:peptide deformylase; D015032:Zinc