The preparation and isolation of carrier-free radio-iodotyrosine

Hadi, Usama A.H.; Malcolme-Lawes, David J.; Oldham, G.

doi:10.1016/0020-708x(77)90103-x

Cited by 9 publications

(3 citation statements)

References 5 publications

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“…First, previous attempts to incorporate either monoiodotyrosine or diiodotyrosine biosynthetically into proteins via in vitro rabbit reticulocyte lysate have failed when these residues are introduced into the system as the free acids (Scherberg et al, 1978). Secondly, this radiolabeled residue is easily synthesized with a specific activity within detectable levels for cell-free translation with rabbit reticulocyte lysate (Hadi et al, 1977). Scheme I: Synthesis" of Acylated Dinucleotide 9 HNR4 7:…”

Section: Resultsmentioning

confidence: 99%

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Site-specific incorporation of nonnatural residues during in vitro protein biosynthesis with semi-synthetic aminoacyl-tRNAs

Bain

Diala²,

Glabe³

et al. 1991

Biochemistry

105

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A method is presented for the incorporation of nonnatural amino acids into proteins during in vitro cell-free translation. A combination of chemical synthesis and run-off transcription was employed to prepare a semisynthetic, nonhypermodified tRNA(Gly) nonsense suppressor acylated with L-3-[125I]iodotyrosine. The presence of this synthetic tRNA during in vitro translation of mRNA containing a nonsense suppression site (e.g., a UAG termination codon) results in the incorporation of the nonnatural amino acid L-3-iodotyrosine into the polypeptide exclusively at the position corresponding to that site. Incorporation of the nonnatural amino acid L-3-[125I]iodotyrosine into the model polypeptide was assessed by quantitative and unambiguous determination of suppression efficiency, read-through, and site specificity of incorporation. Minor modifications of the method employed in this initial experiment also allow the rapid analysis of unlabeled acylated tRNA analogues. Under optimum conditions, the unlabeled amino acid L-3-iodotyrosine was found to be incorporated with a suppression efficiency of 65%. Other nonnatural residues, including N-methylphenylalanine, D-phenylalanine, and phenyllactic acid, were tested in the assay under these same conditions. Suppression efficiencies for this series ranged from 0 to 72% depending on the structure of the residue incorporated. Several other aspects of this methodology, such as tRNA structure and context effects, are briefly discussed.

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Section: Resultsmentioning

confidence: 99%

“…Carrier-free Na 125I (-2500 Ci/mmol) was obtained from ICN Biomedicals, Inc., and l-[35S]methionine (>800 Ci/mmol) and L-[3,4,5-3H]leucine (143 Ci/mmol) were purchased from New England Nuclear Research Products. Carrier-free l-3-[125I]iodotyrosine was prepared by the method of Hadi et al (1977). General Methods.…”

Section: Methodsmentioning

confidence: 99%

Site-specific incorporation of nonnatural residues during in vitro protein biosynthesis with semi-synthetic aminoacyl-tRNAs

Bain

Diala²,

Glabe³

et al. 1991

Biochemistry

105

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show abstract

“…Order to obtain pure labelled molecules without electrolytes [19], water was used as eluent. The disadvantage of using pure water as eluent is the large elution volume, as can be seen for example in Table 1 for the 5-bromo-2'-deoxyuridine mixture.…”

Section: Methodsmentioning

confidence: 99%

On the Preparation of⁸²Br-Biomolecules. A Direct Bromination via Neutron Irradiated Bromates

Alfassi

1979

Radiochimica Acta

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In Vitro Dehalogenation of para-Substituted Aromatic Halides in Rat Liver Preparations

Scott

Sinsheimer

1984

Journal of Pharmaceutical Sciences

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The preparation and isolation of carrier-free radio-iodotyrosine

Cited by 9 publications

References 5 publications

Site-specific incorporation of nonnatural residues during in vitro protein biosynthesis with semi-synthetic aminoacyl-tRNAs

Site-specific incorporation of nonnatural residues during in vitro protein biosynthesis with semi-synthetic aminoacyl-tRNAs

On the Preparation of⁸²Br-Biomolecules. A Direct Bromination via Neutron Irradiated Bromates

In Vitro Dehalogenation of para-Substituted Aromatic Halides in Rat Liver Preparations

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The preparation and isolation of carrier-free radio-iodotyrosine

Cited by 9 publications

References 5 publications

Site-specific incorporation of nonnatural residues during in vitro protein biosynthesis with semi-synthetic aminoacyl-tRNAs

Site-specific incorporation of nonnatural residues during in vitro protein biosynthesis with semi-synthetic aminoacyl-tRNAs

On the Preparation of82Br-Biomolecules. A Direct Bromination via Neutron Irradiated Bromates

In Vitro Dehalogenation of para-Substituted Aromatic Halides in Rat Liver Preparations

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On the Preparation of⁸²Br-Biomolecules. A Direct Bromination via Neutron Irradiated Bromates