In vivo protein labeling with trimethoprim conjugates: A flexible chemical tag

Lawrence W. Miller; Yunfei Cai; Michael P. Sheetz; Virginia W. Cornish

doi:10.1038/nmeth749

In vivo protein labeling with trimethoprim conjugates: A flexible chemical tag

Lawrence W. Miller, Yunfei Cai, Michael P. Sheetz, Virginia W. Cornish

Research output: Contribution to journal › Article › peer-review

260 Scopus citations

Abstract

The introduction of green fluorescent protein and its variants (GFPs) has allowed protein analysis at the level of the cell. Now, chemical methods are needed to label proteins in vivo with a wider variety of functionalities so that mechanistic questions about protein function in the complex cellular environment can be addressed. Here we demonstrate that trimethoprim derivatives can be used to selectively tag Escherichia coli dihydrofolate reductase (eDHFR) fusion proteins in wild-type mammalian cells with minimal background and fast kinetics.

Original language	English (US)
Pages (from-to)	255-257
Number of pages	3
Journal	Nature Methods
Volume	2
Issue number	4
DOIs	https://doi.org/10.1038/nmeth749
State	Published - Apr 2005
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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title = "In vivo protein labeling with trimethoprim conjugates: A flexible chemical tag",

abstract = "The introduction of green fluorescent protein and its variants (GFPs) has allowed protein analysis at the level of the cell. Now, chemical methods are needed to label proteins in vivo with a wider variety of functionalities so that mechanistic questions about protein function in the complex cellular environment can be addressed. Here we demonstrate that trimethoprim derivatives can be used to selectively tag Escherichia coli dihydrofolate reductase (eDHFR) fusion proteins in wild-type mammalian cells with minimal background and fast kinetics.",

author = "Miller, {Lawrence W.} and Yunfei Cai and Sheetz, {Michael P.} and Cornish, {Virginia W.}",

note = "Funding Information: This research was supported by the National Institutes of Health (GM071754-01). V.W.C. is a recipient of a Beckman Young Investigator Award, a Burroughs Wellcome Fund New Investigator Award in the Toxicological Sciences, a Camille and Henry Dreyfus New Faculty Award and a National Science Foundation Career Award.",

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AU - Cai, Yunfei

AU - Sheetz, Michael P.

AU - Cornish, Virginia W.

N1 - Funding Information: This research was supported by the National Institutes of Health (GM071754-01). V.W.C. is a recipient of a Beckman Young Investigator Award, a Burroughs Wellcome Fund New Investigator Award in the Toxicological Sciences, a Camille and Henry Dreyfus New Faculty Award and a National Science Foundation Career Award.

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AB - The introduction of green fluorescent protein and its variants (GFPs) has allowed protein analysis at the level of the cell. Now, chemical methods are needed to label proteins in vivo with a wider variety of functionalities so that mechanistic questions about protein function in the complex cellular environment can be addressed. Here we demonstrate that trimethoprim derivatives can be used to selectively tag Escherichia coli dihydrofolate reductase (eDHFR) fusion proteins in wild-type mammalian cells with minimal background and fast kinetics.

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In vivo protein labeling with trimethoprim conjugates: A flexible chemical tag

Abstract

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