• YAKHAK HOEJI
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• v.47 no.3
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• pp.184-189
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• 2003

Peptide deformylase (PDF) is essential and unique to bacteria, thus making it an attractive target for the discovery of novel antibacterial drugs. PDF deformylates the N-formylmethionine of newly synthesized polypeptides in prokaryotes. In this study, a pdf gene from Staphylococcus aureus 6538p was cloned in pET-14b vector and PDF protein was over-produced in Escherichia coli BL21 (DE3). NH$_2$-terminal His-tagged PDF protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography. Enzymatic activity of purified 6xHis-tagged PDF was tested on the substrate (formyl-Methionine-Alanine-Serine) by formate dehydrogenase-coupled spectrometric assay of peptide deformylase. For the discovery of new PDF inhibitors from chemical libraries and culture broths of soil bacteria, a target-oriented screening system using a 96-well plate was developed. About 3,000 commercial chemical libraries were tested in this screening system, and 2 chemicals (0.07％) among them showed an inhibitory activity against PDF enzyme. This result showed that a new screening system can be used for the discovery of new PDF inhibitors.

• BMB Reports
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• v.40 no.6
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• pp.1050-1057
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• 2007

Peptide deformylase (PDF) is a metalloenzyme that removes the N-terminal formyl groups from newly synthesized proteins. It is essential for bacterial survival, and is therefore-considered as a potential target for antimicrobial chemotherapy. However, some bacteria including medically relevant pathogens possess two or more def-like genes. Here we have examined two PDFs from Bacillus cereus. The two share only 32% sequence identity and the crystal structures show overall similarity with PDF2 having a longer C-terminus. However, there are differences at the two active sites, and these differences appear to contribute to the activity difference seen between the two. BcPDF2 is found as a dimer in the crystal form with two additional actinonin bound at that interface.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.163.4-164
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• 2003

Peptide deformylase (PDF) is essential and unique to bacteria for cytoplasmic protein synthesis, but not required in eukaryotes, thus making it an attractive target for the discovery of novel antibacterial drugs. Protein synthesis in eubacteria, under normal conditions, is initiated by formyl-methionyl-tRNA. PDF removes the formyl-group of N- formylmethionine of newly synthesized polypeptides to produce a mature protein. In this study, a pdf gene from Staphylococcus aureus 6538p was cloned in pET-14b vector and transformed in Escherichia coli BL21 (DE3). (omitted)