The Selective Incorporation of Alkenes into Proteins in
            <i>Escherichia coli</i>

Zhang, Zhiwen; Wang, Lei; Brock, Ansgar; Schultz, Peter G.

doi:10.1002/1521-3773(20020802)41:15<2840::aid-anie2840>3.0.co;2-#

Cited by 86 publications

(43 citation statements)

References 21 publications

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“…To date, Ͼ25 unnatural amino acids have been incorporated into proteins in response to the amber nonsense codon, TAG, with high translational efficiency and fidelity. These include amino acids that can be photocrosslinked, glycosylated amino acids, chemically reactive amino acids (containing keto, alkene, or alkyne groups), redox active amino acids and fluorescent amino acids (2,3,5,(7)(8)(9). To genetically encode 22 or more amino acids requires additional codons that uniquely specify each new amino acid.…”

mentioning

confidence: 99%

An expanded genetic code with a functional quadruplet codon

Anderson

Santoro

et al. 2004

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

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305

251

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With few exceptions the genetic codes of all known organisms encode the same 20 amino acids, yet all that is required to add a new building block are a unique tRNA͞aminoacyl-tRNA synthetase pair, a source of the amino acid, and a unique codon that specifies the amino acid. For example, the amber nonsense codon, TAG, together with orthogonal Methanococcus jannaschii or Escherichia coli tRNA͞synthetase pairs have been used to genetically encode a variety of unnatural amino acids in E. coli and yeast, respectively. However, the availability of noncoding triplet codons ultimately limits the number of amino acids encoded by any organism. Here, we report the design and generation of an orthogonal synthetase͞ tRNA pair derived from archaeal tRNA Lys sequences that efficiently and selectively incorporates an unnatural amino acid into proteins in response to the quadruplet codon, AGGA. Frameshift suppression with L-homoglutamine (hGln) does not significantly affect protein yields or cell growth rates and is mutually orthogonal with amber suppression, permitting the simultaneous incorporation of two unnatural amino acids, hGln and O-methyl-L-tyrosine, at distinct positions within myoglobin. This work suggests that neither the number of available triplet codons nor the translational machinery itself represents a significant barrier to further expansion of the genetic code.

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mentioning

confidence: 99%

An expanded genetic code with a functional quadruplet codon

Anderson

Santoro

et al. 2004

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

Self Cite

305

251

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“…In this approach, an orthogonal transfer RNA (tRNA)-aminoacyl tRNA synthetase pair is evolved that uniquely recognizes the amino acid of interest and selectively incorporates it into proteins in response to the amber nonsense codon, TAG. This methodology has been used to site-specifically incorporate a variety of unnatural amino acids into proteins with high fidelity and good efficiency, including amino acids with novel functional groups (3)(4)(5)(6), photocrosslinkers (7,8), heavy atoms, sugars (9), and redox active moieties. In addition, new orthogonal tRNA-synthetase pairs have been evolved from leucyl (10), lysyl, glutaminyl (11), aspartyl (12), and tyrosyl (13) tRNA-synthetase pairs to expand the number and structural diversity of amino acids that can be genetically encoded in bacteria and yeast.…”

mentioning

confidence: 99%

Selective incorporation of 5-hydroxytryptophan into proteins in mammalian cells

Zhang

Alfonta

Tian

et al. 2004

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

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“…2 (6-8)) [33, [62][63][64]. Alkenes can undergo olefin metathesis reactions with another alkene molecule [64,65], or Mizoroki-Heck reactions with unsaturated halides [66,67].…”

Section: Bioorthogonal Labeling Of Proteinsmentioning

confidence: 99%

“…Both need appropriate organometallic catalysts, however. Recent research has reported water-soluble catalysts, but their applications to diverse proteins and in living cells remain to be established [60][61][62][63]. In addition, Lin and coworkers reported a UV light-induced coupling reaction between olefins and tetrazoles (Fig.…”

Section: Bioorthogonal Labeling Of Proteinsmentioning

confidence: 99%

Biochemical analysis with the expanded genetic lexicon

2012

Anal Bioanal Chem

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The information used to build proteins is stored in the genetic material of every organism. In nature, ribosomes use 20 native amino acids to synthesize proteins in most circumstances. However, laboratory efforts to expand the genetic repertoire of living cells and organisms have successfully encoded more than 80 nonnative amino acids in E. coli, yeast, and other eukaryotic systems. The selectivity, fidelity, and site-specificity provided by the technology have enabled unprecedented flexibility in manipulating protein sequences and functions in cells. Various biophysical probes can be chemically conjugated or directly incorporated at specific residues in proteins, and corresponding analytical techniques can then be used to answer diverse biological questions. This review summarizes the methodology of genetic code expansion and its recent progress, and discusses the applications of commonly used analytical methods.

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The Selective Incorporation of Alkenes into Proteins in Escherichia coli

Cited by 86 publications

References 21 publications

An expanded genetic code with a functional quadruplet codon

An expanded genetic code with a functional quadruplet codon

Selective incorporation of 5-hydroxytryptophan into proteins in mammalian cells

Biochemical analysis with the expanded genetic lexicon

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