Non-natural labeled amino acid and method for producing a conjugate of said amino acid and trna
a technology conjugates, which is applied in the field of non-natural amino acids, can solve the problems of disadvantageous low yield, difficult to achieve favorable results in respect of the reproducibility of the modification site and/or the regulation of the number of modifications, and non-natural amino acids cannot be introduced to a protein molecule. , to achieve the effect of easy separation and purification, and easy synthesizing
Inactive Publication Date: 2008-09-11
PROTEIN EXPRESS +1
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AI Technical Summary
Benefits of technology
This approach allows for the synthesis of functional proteins with desired fluorescent properties, enhancing detection and analysis capabilities in various fields by ensuring high reproducibility and specificity of label incorporation.
Problems solved by technology
On the contrary, a favorable outcome is hard to attain in respect of the reproducibility of the modification site and / or the regulation of the number of modifications.
In this method that utilizes a termination codon, however, two or more kinds of non-natural amino acids cannot be introduced to a protein molecule, and the yield is disadvantageously low.
However, an amino acid having a side chain with a bulky molecular structure such as l-pyrenylalanine or ferrocenylalanine cannot be introduced to a protein through the natural protein synthesis system.
When these fluorescent substances are used as label compounds for non-natural amino acids, however, it is difficult to introduce them to the proteins by a method that utilizes a protein synthesis system due to their large molecular weights.
The possibility of a label compound being introduced to a protein is disadvantageously limited by the method that utilizes a protein synthesis system due to its molecular weight.
Method used
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Embodiment Construction
[0081]The present invention is hereafter described in greater detail with reference to the following examples, although the present invention is not limited thereto.
Synthesis of BODIPY FL-AF-pdCpA, BODIPY FL-X-AF-pdCpA, BODIPY FL-K-pdCpA, and BODIPY FL-X-K-pdCpA
Synthesis of AF-pdCpA
[0082]di-Boc-aminophenylalanine cyanomethyl ester (1.1 μmol) was added to 5 μl of a solution of pdCpA tetra-n-butylammonium (0.044 μmol / μl) in DMF, and the mixture was allowed to react at room temperature for 2 hours. After the reaction progress was confirmed by HPLC, 1.4 ml of diethyl ether was added to the reaction solution. The resultant was agitated and centrifuged at 15,000 rpm for 5 minutes, and the supernatant was removed. The precipitate was dissolved by adding 20 μl of acetonitrile, 1.4 ml of diethyl ether was added thereto, and the resultant was agitated, followed by centrifugation at 15,000 rpm for 5 minutes. The supernatant was removed, the precipitate was dissolved by adding 20 μl of acetonit...
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This invention relates to: a labeled amino acid that can be introduced into a protein with the aid of a protein synthesis system; a functional protein having functions derived from a label compound; a labeled amino acid comprising an aromatic ring bound to an amino acid side chain and a label compound bound thereto via the aromatic ring; a functional protein to which the labeled amino acid has been introduced; and a novel method for effectively obtaining a labeled amino acid-tRNA complex.
Description
CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation application of prior U.S. patent application Ser. No. 10 / 490,508, which was the National Stage of International Application No. PCT / JP03 / 08970, filed Jul. 15, 2003. The parent application claims priority to Japanese Application No. 2002-209736, filed Jul. 18, 2002, the disclosures of which are incorporated herein by reference in their entireties.BACKGROUND OF THE INVENTION[0002]1. Technical Field[0003]The present invention provides, as a non-natural amino acid, a labeled amino acid in which a label compound is bound directly or via a spacer to an amino acid having an amino acid skeleton with an aromatic ring such as a benzene ring on its side chain. This labeled amino acid is introduced to a protein by utilizing a protein synthesis system, thereby providing a functional protein having functions derived from the label compound with high reproducibility. Further, the present invention relates to a novel me...
Claims
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IPC IPC(8): C07K16/40C12N15/13C07C229/52C12P21/06C07K14/435C07F5/02C09B57/00C09B57/10
CPCC07B59/001C07F5/022C12P21/02C09B57/00C09B57/10C07F5/027
InventorHOHSAKA, TAKAHIROSISIDO, MASAHIKO
OwnerPROTEIN EXPRESS