Drug target validation: Lethal infection blocked by inducible peptide

Tao, Jianshi; Wendler, Philip A.; Connelly, Gene; Lim, Audrey; Zhang, Jiansu; King, Megan; Li, Tongchuan; Silverman, Jared; Schimmel, Paul; Tally, Francis P.

doi:10.1073/pnas.97.2.783

Cited by 50 publications

(24 citation statements)

References 14 publications

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“…18) have successfully been used in yeast cells, E. coli, Drosophila, and cultured human cells. They are a useful tool for basic research, but they also represent a new approach in drug target validation and drug discovery (18,19,(31)(32)(33)(34). The application of dominant negative peptide aptamers to inhibit protein functions in vivo in a living multicellular organism has been reported so far only for Drosophila (19,35).…”

Section: Discussionmentioning

confidence: 99%

Peptide-mediated broad-spectrum plant resistance to tospoviruses

Rudolph

Schreier

Uhrig

2003

Proc. Natl. Acad. Sci. U.S.A.

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Plant viruses have a significant impact on agronomic losses worldwide. A new strategy for engineering virus-resistant plants by transgenic expression of a dominant interfering peptide is presented here. This peptide of 29 aa strongly interacts with the nucleocapsid proteins (N) of different tospoviruses. Transgenic Nicotiana benthamiana lines expressing the peptide fused to a carrier protein were challenged with five different tospoviruses that have a nucleocapsid protein interacting with the peptide. In the transgenic plants, strong resistance to tomato spotted wilt virus, tomato chlorotic spot virus, groundnut ring spot virus, and chrysanthemum stem necrosis virus was observed. This therefore demonstrates the feasibility of using peptide ''aptamers'' as an in vivo tool to control viral infection in higher plants.

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Section: Discussionmentioning

confidence: 99%

Peptide-mediated broad-spectrum plant resistance to tospoviruses

Rudolph

Schreier

Uhrig

2003

Proc. Natl. Acad. Sci. U.S.A.

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“…Peptides were synthesized to include the 12-to 15-amino-acid sequence displayed by the phage followed by a short linker and a biotin at the C terminus to allow labeling for analysis of peptide binding. Isolation and use of ProRS-specific phage has been described previously (24,46); peptide Pro-3 in those publications is referred to as peptide 72 in this work.…”

mentioning

confidence: 99%

“…In an extension of our previous work in isolating a peptide inhibitor of prolyl-tRNA synthetase (ProRS) (24), Tao et al showed that intracellular expression of this peptide fused to GST could slow bacterial growth and rescue mice from an otherwise lethal dose of virulent Escherichia coli. (46). Specific inhibition of ProRS in the cell was demonstrated by a decrease in the intracellular pool of charged tRNA Pro .…”

mentioning

confidence: 99%

Intracellular Expression of Peptide Fusions for Demonstration of Protein Essentiality in Bacteria

Benson¹,

Gottlin²,

Christensen³

et al. 2003

Antimicrob Agents Chemother

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We describe a "protein knockout" technique that can be used to identify essential proteins in bacteria. This technique uses phage display to select peptides that bind specifically to purified target proteins. The peptides are expressed intracellularly and cause inhibition of growth when the protein is essential. In this study, peptides that each specifically bind to one of seven essential proteins were identified by phage display and then expressed as fusions to glutathione S-transferase in Escherichia coli. Expression of peptide fusions directed against E. coli DnaN, LpxA, RpoD, ProRS, SecA, GyrA, and Era each dramatically inhibited cell growth. Under the same conditions, a fusion with a randomized peptide sequence did not inhibit cell growth. In growthinhibited cells, inhibition could be relieved by concurrent overexpression of the relevant target protein but not by coexpression of an irrelevant protein, indicating that growth inhibition was due to a specific interaction of the expressed peptide with its target. The protein knockout technique can be used to assess the essentiality of genes of unknown function emerging from the sequencing of microbial genomes. This technique can also be used to validate proteins as drug targets, and their corresponding peptides as screening tools, for discovery of new antimicrobial agents.

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“…Information obtained from peptide library approaches also provides useful information and tools for drug discovery. For example, binding information derived from phage display methods has been used to validate particular proteins as potential drug targets in vivo (Stauffer et al, 1997;Tao et al, 2000). In addition, phage display ligands have been used to guide the design of peptidomimetics and for development of high-throughput small molecule inhibitor screens (Kay et al, 1998;Gron et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

Exploring Biochemistry and Cellular Biology with Protein Libraries

Diaz

Howard²,

Neubauer³

et al. 2003

Current Issues in Molecular Biology

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Polypeptide libraries cast a broad net for defining enzyme and binding protein specificities. In addition to uncovering rules for molecular recognition, the binding preferences and functional group tolerances from such libraries can reveal mechanisms underlying biochemical and cellular processes. Ligands obtained from protein libraries can also provide pharmaceutical lead compounds and even reagents to further explore cell biology. Here, we review selected recent examples of protein libraries demonstrating these principles. In particular, we focus on combinatorial libraries composed of randomized peptides or variations of a single protein. The characteristics of various techniques for library constructions and screening are also briefly surveyed.

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Drug target validation: Lethal infection blocked by inducible peptide

Cited by 50 publications

References 14 publications

Peptide-mediated broad-spectrum plant resistance to tospoviruses

Peptide-mediated broad-spectrum plant resistance to tospoviruses

Intracellular Expression of Peptide Fusions for Demonstration of Protein Essentiality in Bacteria

Exploring Biochemistry and Cellular Biology with Protein Libraries

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