Back to Search
Start Over
Purification, characterization, and inhibition of peptide deformylase from Escherichia coli
- Source :
- Biochemistry. 36(45)
- Publication Year :
- 1997
-
Abstract
- Peptide deformylase (EC 3.5.1.31) catalyzes the removal of a formyl group from the N-termini of nascent ribosome-synthesized polypeptides, an obligatory step during protein maturation in eubacteria. Since its discovery in crude Escherichia coli extracts 3 decades ago, the deformylase has resisted all attempts of purification or characterization due to its extraordinary lability. By placing the coding sequence (def gene) of Escherichia coli deformylase behind a bacteriophage T7 promoter, we have, however, been able to overexpress this deformylase in Escherichia coli. Overproduction has allowed the purification of50 mg of deformylase enzyme from each liter of cell culture. Purified deformylase is highly active toward N-formylated peptide substrates. A new, sensitive assay for the deformylase has been developed by measuring the amount of released formate using a formate dehydrogenase. This has allowed for the assessment of the catalytic properties of peptide deformylase using a series of synthetic N-formylated peptides as substrates. The deformylase exhibits strong preference for an L-methionine or the isosteric norleucine at the N-terminus of a substrate and has broad specificity for the rest of the residues. Small divalent metal chelators strongly inhibit the E. coli deformylase. In particular, certain 1,2- and 1,3-dithiol compounds act as potent, time-dependent inhibitors of the peptide deformylase.
- Subjects :
- Cations, Divalent
Peptide
Biology
medicine.disease_cause
Formate dehydrogenase
Biochemistry
Aminopeptidases
Amidohydrolases
Substrate Specificity
Bacteriophage
Peptide deformylase
Enzyme Stability
Metalloproteins
medicine
Escherichia coli
Amino Acid Sequence
Sulfhydryl Compounds
Overproduction
Protein maturation
Chelating Agents
chemistry.chemical_classification
Hydrogen-Ion Concentration
biology.organism_classification
Recombinant Proteins
Kinetics
Enzyme
chemistry
Subjects
Details
- ISSN :
- 00062960
- Volume :
- 36
- Issue :
- 45
- Database :
- OpenAIRE
- Journal :
- Biochemistry
- Accession number :
- edsair.doi.dedup.....ad4153f619b0d37a01c077638e09e85f