PMID:15110195

Citation	Nguyen, KT, Hu, X and Pei, D (2004) Slow-binding inhibition of peptide deformylase by cyclic peptidomimetics as revealed by a new spectrophotometric assay. Bioorg. Chem. 32:178-91
Abstract	A new spectrophotometric/fluorimetric assay for peptide deformylase (PDF) has been developed by coupling the PDF reaction with that of dipeptidyl peptidase I (DPPI) and using N-formyl-Met-Lys-AMC as substrate. Removal of the N-terminal formyl group by PDF renders the dipeptide an efficient substrate of DPPI, which subsequently removes the dipeptidyl units to release 7-amino-4-methylcoumarin as the chromophore/fluorophore. The PDF reaction is conveniently monitored on a UV-Vis spectrophotometer or a fluorimeter in a continuous fashion. The utility of the assay was demonstrated by determining the catalytic activity of PDF and the inhibition constants of PDF inhibitors. These studies revealed the slow-binding behavior of a previously reported macrocyclic PDF inhibitor. This method offers several advantages over the existing PDF assays and should be particularly useful for screening PDF inhibitors in the continuous fashion.
Links	PubMed Online version:10.1016/j.bioorg.2004.01.001
Keywords	Amidohydrolases/antagonists & inhibitors; Amidohydrolases/metabolism; Biomimetic Materials/chemistry; Cathepsin C/chemistry; Cathepsin C/metabolism; Enzyme Inhibitors/chemistry; Enzyme Inhibitors/pharmacology; Escherichia coli/enzymology; Kinetics; Peptides, Cyclic/pharmacology; Spectrometry, Fluorescence/methods; Spectrophotometry, Ultraviolet/methods; Substrate Specificity
edit table

Significance

You can help EcoliWiki by summarizing why this paper is useful

Useful Materials and Methods

You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.

Annotations

<annotationlinks/>

EcoliWiki Links

Add links to pages that link here (e.g. gene, product, method pages)

References

See Help:References for how to manage references in EcoliWiki.