Preparation of Alloproteins Endowed with Artificial Functions

赋予人工功能的同种异体蛋白的制备

• 批准号: 06403034
• 负责人: SISIDO Masahiko
• 金额: $ 16.83万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06403034/
• 关键词: Alloproteins; Nonnatural Amino Acids; Extension of Genetic Code; Antibodies; Electron Transfer; たんぱく質; ストレプトアビジン; エネルギー移動; フレームシフト; 蛍光スペクトル; 超たんぱく質; 人工機能; ストップコドン; ストレプアビジン; 蛍光性アミノ酸; 核酸塩基アミノ酸

Proteins containing nonnatural amino acids at specific sites (alloproteins) were preparaed by the extension of gentic engineering technique. The alloproteins were endowed with artificial functions by the incorporation of nonnatural amino acids carrying specialty side groups. The results are summarized as follows.(1)Extension of Amino Acids : A variety of nonnatural amino acids carrying specialty side groups were newly synthesized and tested for the adaptability to in vitro E.Coli ribosomal system. Those that carry linear aromatic groups were found to be accepted by the ribosomal system. The results opened a way for the photo-and electro-functionalization of proteins.(2)Extension of Genetic Code : Codons that may be assigned to nonnatural amino acids were searched. Some 4-base codons, such as CGGG and AGGU were found to be effective for this purpose. The 4-base codons may be used for multiple incorporation of different nonnatural amino acids into single proteins(3)Use of Monoclonal Antibodies as the Controlling Factor of Alloproteins : Monoclonal antibodies aganinst nonnatural amino acids were prepared. Those were used to photo-and electro-contorl of alloproteins that carry azobenzene groups or anthraquinonyl groups.

通过基因工程技术的延伸，在特定位点含有非天然氨基酸的蛋白质（异源蛋白）被克隆出来。通过掺入带有特殊侧基的非天然氨基酸，赋予异源蛋白人工功能。结果总结如下。（1）氨基酸的延伸：新合成了多种带有特殊侧基的非天然氨基酸，并测试了它们对体外大肠杆菌核糖体系统的适应性。那些携带线性芳香基团的被发现被核糖体系统所接受。该结果为蛋白质的光电功能化开辟了一条道路。（2）遗传密码的延伸：搜索可能分配给非天然氨基酸的密码子。一些4碱基密码子，如CGGG和AGGU被发现是有效的，用于此目的。4碱基密码子可用于将不同的非天然氨基酸多重掺入单个蛋白质中。（3）单克隆抗体作为同种蛋白的控制因子：制备了抗非天然氨基酸的单克隆抗体。它们被用来对带有偶氮苯基或蒽醌基的同种蛋白进行光电控制。

项目成果

M.Imaizumi et al: "Fluorescence Behavior of Pyrenyl Groups Bound to Anti-Pyrenyl Antibody" Journal of Physical Chemistry. 99・11. 3810-3814 (1995)

M.Imaizumi 等：“与抗芘基抗体结合的芘基团的荧光行为”物理化学杂志 99・11 3810-3814（1995）。

宍戸昌彦,芳坂貴弘: "機能性非天然アミノ酸を含む蛋白質の合成と応用" 化学と工業. 49(4). 515-517 (1996)

Masahiko Shishido，Takahiro Yoshisaka：“含有功能性非天然氨基酸的蛋白质的合成和应用” Kagaku to Kogyo 49(4) 515-517 (1996)。

M.Harada: "Photocontrolled Uptake and Release of Photochromic Heptens by Monoclonal Antibodies" Bulletin of the Chemical Society,Japan. 67. 1380-1385 (1994)

M.Harada：“单克隆抗体对光致变色庚烯的光控摄取和释放”日本化学会公报。

M.Imaizumi: "Fluorescence Behavior of Pyrenyl Groups bound to Anti-Pyrenyl Antibody" Journal of Physical Chemistry. (印刷中).

M.Imaizumi：“与抗芘基抗体结合的芘基团的荧光行为”物理化学杂志（正在出版）。

宍戸 昌彦 他: "機能性非天然アミノ酸を含む蛋白質の合成と応用" 化学と工業. 49・4. 515-517 (1996)

Masahiko Shishido等人：“含有功能性非天然氨基酸的蛋白质的合成和应用”化学与工业49・4（1996）。