Title

Characterization of a human peptide deformylase: Implications for antibacterial drug design

Authors

Authors

K. T. Nguyen; X. B. Hu; C. Colton; R. Chakrabarti; M. X. Zhu;D. H. Pei

Comments

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Abbreviated Journal Title

Biochemistry

Keywords

ESCHERICHIA-COLI; SPECTROPHOTOMETRIC ASSAY; PROTEIN; AGENTS; IRON; FORMYLTRANSFERASE; FORMYLMETHIONINE; IDENTIFICATION; PURIFICATION; TRANSLATION; Biochemistry & Molecular Biology

Abstract

Ribosomal protein synthesis in eubacteria and eukaryotic organelles initiates with an N-formylmethionyl-tRNA(i), resulting in N-terminal formylation of all nascent polypeptides. Peptide deformylase (PDF) catalyzes the subsequent removal of the N-terminal formyl group from the majority of bacterial proteins. Deformylation was for a long time thought to be a feature unique to the prokaryotes, making PDF an attractive target for designing novel antibiotics. However, recent genomic sequencing has revealed PDF-like sequences in many eukaryotes, including man. In this work, the cDNA encoding Homo sapiens PDF (HsPDF) has been cloned and a truncated form that lacks the N-terminal 58-amino-acid targeting sequence was overexpressed in Escherichia coli. The recombinant, Co2+-substituted protein is catalytically active in deformylating N-formylated peptides, shares many of the properties of bacterial PDF, and is strongly inhibited by specific PDF inhibitors. Expression of HsPDF fused to the enhanced green fluorescence protein in human embryonic kidney cells revealed its location in the mitochondrion. However, HsPDF is much less active than its bacterial counterpart, providing a possible explanation for the apparent lack of deformylation in the mammalian mitochondria. The lower catalytic activity is at least partially due to mutation of a highly conserved residue (Leu-91 in E. coli PDF) in mammalian PDF. PDF inhibitors had no detectable effect on two different human cell lines. These results suggest that HsPDF is likely an evolutional remnant without any functional role in protein formylation/deformylation and validates PDF as an excellent target for antibacterial drug design.

Journal Title

Biochemistry

Volume

42

Issue/Number

33

Publication Date

1-1-2003

Document Type

Article

Language

English

First Page

9952

Last Page

9958

WOS Identifier

WOS:000184863500013

ISSN

0006-2960

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