X-Ray Crystal Structure Analysis of Recombinant Interleukin-2 and beta-Interferon

重组白细胞介素2和β-干扰素的X射线晶体结构分析

• 批准号: 62490005
• 负责人: MITSUI Yukio
• 金额: $ 4.03万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62490005/
• 关键词: X-ray structure analysis; Interleukin-2; beta-Interferon; Interferon; Lymphokine; Cytokine; 遺伝子工学; β-インターフェロン; リンフォカイン

1) Thanks to recombinant DNA technique, certain lymphokines and cytokines such as interleukin-2 (IL-2) and beta-Interferon (IFN-beta) are now available with the amount of sample feasible for crystallographic study. However the crystals obtained from such sample tend to be disordered, twinned and brittle. To cope with such setbacks, we first improved the method of X-ray diffraction data collection by employing synchrotron beam, imaging plate and by improving the softwares.2) We succeeded in growing hexagonal, orthorhombic and monoclimic crystals of IL-2. However even the best of these (the monoclinic crystal) showed marked weakness against radiation damage. Although we were able to show that sulfoxidation of Met 104 was responsible, we could not find effective remedy for this effect.3) We succeeded in growing orthorhombic and hexagonal crystals of IFN-beta. The hexagonal crystal seems almost ideal for full scale structure analysis. Three promising heavy-atom derivatives were found. Attempt at locating the heavy-atom binding sites are under way. We expect to get three-dimensional structure of IFN-beta within a year.

1)由于重组DNA技术，某些淋巴因子和细胞因子，如白细胞介素-2（IL-2）和β-干扰素（IFN-β），现在可以获得晶体学研究可行的样品量。然而，从这样的样品获得的晶体往往是无序的，孪晶和脆性。为了科普这些困难，我们首先通过同步辐射、成像板和改进软件改进了X射线衍射数据采集的方法。2）成功地生长了六方、正交和单斜晶体IL-2。然而，即使是其中最好的（单斜晶体）也显示出对辐射损伤的明显弱点。虽然我们能够证明Met 104的磺基氧化是负责的，但我们无法找到有效的补救措施。3）我们成功地生长了IFN-β的正交和六方晶体。六方晶体似乎几乎是理想的全尺寸结构分析。发现了三个有希望的重原子衍生物，并正在尝试定位重原子结合位点。我们希望在一年内得到IFN-β的三维结构。

项目成果

松田進,千田俊哉,三井幸雄: J.Biol,Chem. (1989)

Susumu Matsuda、Toshiya Senda、Yukio Mitsui：J.Biol，Chem。

C.Sano;K.Ishikawa;Y.Mitsui;Y.Iitaka: J.Biol.Chem.262. 4766-4769 (1987)

C.Sano；K.Ishikawa；Y.Mitsui；Y.Iitaka：J.Biol.Chem.262。

平山令明,三井幸雄: "化学増刊113プロテイン・エンジニアリング1.2章 X線結晶解析" 化学同人, 280 (1988)

Reiaki Hirayama、Yukio Mitsui：“化学特别版 113 蛋白质工程第 1.2 章 X 射线晶体分析” Kagaku Doujin，280（1988）

S.Matsuda; T.Senda; Y.Mitsui: "New Crystal Form of Murine recombinant Interferon-" J. Biol. Chem.(1989)

松田S.

佐野千明,石川好紀,三井幸雄: J.Biol Chem.262. 4766-4769 (1987)

Chiaki Sano、Yoshiki Ishikawa、Yukio Mitsui：J.Biol Chem.262 4766-4769 (1987)。