Peptide deformylase as an antibacterial drug target:: Assays for detection of its inhibition in Escherichia coli cell homogenates and intact cells

被引：15

作者：

Apfel, CM ^{[1
]}

Evers, S ^{[1
]}

Hubschwerlen, C ^{[1
]}

Pirson, W ^{[1
]}

Page, MGP ^{[1
]}

Keck, W ^{[1
]}

机构：

[1] F Hoffmann La Roche & Co Ltd, PRBMH, CH-4070 Basel, Switzerland

来源：

ANTIMICROBIAL AGENTS AND CHEMOTHERAPY | 2001年 / 45卷 / 04期

关键词：

D O I：

10.1128/AAC.45.4.1053-1057.2001

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

An assay was developed to determine the activity of peptide deformylase (PDF) inhibitors under conditions as close as possible to the physiological situation. The assay principle is the detection of N-terminal [S-35]methionine labeling of a protein that contains no internal methionine. If PDF is active, the deformylation of the methionine renders the peptide a substrate for methionine aminopeptidase, resulting in the removal of the N-terminal methionine label. In the presence of a PDF inhibitor, the deformylation is blocked so that the N-formylated peptide is not processed and the label is detected, Using this assay, it is possible to determine the PDF activity under near-physiological conditions in a cell-free transcription-translation system as well as in intact bacterial cells.

引用

页码：1053 / 1057

页数：5

共 19 条

[11] N-FORMYL-METHIONYL-S-RNA [J].

MARCKER, K ;

SANGER, F .

JOURNAL OF MOLECULAR BIOLOGY, 1964, 8 (06) :835-&

[12] GENETIC-CHARACTERIZATION OF POLYPEPTIDE DEFORMYLASE, A DISTINCTIVE ENZYME OF EUBACTERIAL TRANSLATION [J].

MAZEL, D ;

POCHET, S ;

MARLIERE, P .

EMBO JOURNAL, 1994, 13 (04) :914-923

[13] METHIONINE AS TRANSLATION START SIGNAL - A REVIEW OF THE ENZYMES OF THE PATHWAY IN ESCHERICHIA-COLI [J].

MEINNEL, T ;

MECHULAM, Y ;

BLANQUET, S .

BIOCHIMIE, 1993, 75 (12) :1061-1075

[14] CHARACTERIZATION OF THE THERMUS-THERMOPHILUS LOCUS ENCODING PEPTIDE DEFORMYLASE AND METHIONYL-TRNA(F)(MET) FORMYLTRANSFERASE [J].

MEINNEL, T ;

BLANQUET, S .

JOURNAL OF BACTERIOLOGY, 1994, 176 (23) :7387-7390

[15] Formylation is not essential for initiation of protein synthesis in all eubacteria [J].

Newton, DT ;

Creuzenet, C ;

Mangroo, D .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (32) :22143-22146

[16]

SAMBROOK RF, 1989, MOL CLONING LAB MANU

[17] DNA SEQUENCING WITH CHAIN-TERMINATING INHIBITORS [J].

SANGER, F ;

NICKLEN, S ;

COULSON, AR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (12) :5463-5467

[18] Processing of the N termini of nascent polypeptide chains requires deformylation prior to methionine removal [J].

Solbiati, J ;

Chapman-Smith, A ;

Miller, JL ;

Miller, CG ;

Cronan, JE .

JOURNAL OF MOLECULAR BIOLOGY, 1999, 290 (03) :607-614

[19]

STUBER D, 1990, IMMUNOLOGICAL METHOD, V4, P121

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