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    • 3. 发明申请
    • FLUORESCENT ENERGY FOR ELUCIDATING THE 3-D STRUCTURE OF BIOLOGICAL MACROMOLECULES
    • 荧光能量转移用于生物分子的三维结构的教育
    • WO9941607A3
    • 1999-12-09
    • PCT/EP9901008
    • 1999-02-10
    • GMD GMBHHOFFMANN DANIEL
    • HOFFMANN DANIEL
    • C12Q1/68G01N33/533G01N33/58G01N33/68
    • G01N33/6848G01N33/533G01N33/582Y10S435/968Y10S435/973Y10S436/80Y10S436/804Y10S436/805Y10S436/819Y10T436/13
    • The invention relates to a method for the targeted chemical activation of photoactivatable cross-linking molecules around ligand binding pockets and fluorescent groups in macromolecules, especially biological macromolecules, by using fluorescent ligands or fluorescent groups of the macromolecule, and by selecting photoactivatable cross-linking molecules with specific activation energies, to achieve a radiation-free energy transfer (Forster transfer) from the fluorescent ligands or groups to the cross-linking molecules activated thereby. The invention also relates to a method for elucidating the 3-D structure of macromolecules (M), characterized in that a ligand (F) capable of fluorescence with a fluorescence frequency in the range (1 to (2 is introduced into the macromolecule (M) or its physical position to the macromolecule (M) is determined using known methods; one or more photoactivatable bifunctional cross-linking agents (C) with a corresponding excitation frequency in the range of (1 to (2) are bound in a covalent manner between the non-photoactivatable end (S) of the cross-linking agents (C, C', C") and suitable functional groups (m) of the macromolecule (M) in the absence of light; the macromolecule (M) is irradiated above the frequency interval (1 to (2 at a frequency (Q, and by means of a radiation-free transfer (Forster transfer) to neighbouring cross-linking agents (C and/or C"), the photoactivatable end (A and/or A') of the cross-linking element (C and/or C") is activated for reaction with the surface of the macromolecule (M) and reacted with the surface of said macromolecule (M) in accordance with the distance of the ligand (F) capable of fluorescence; the groups linked in pairs are identified using bioanalytical methods, especially specific digestion of the macromolecule (M), the digested fragments are divided, especially by mass, and physical proximities are determined by calculation.
    • 一种用于大分子的三维结构分析(M)的方法,其特征在于,一个引入,或能够在(1范围内的荧光用荧光频率配体(F)至(2(在所述大分子M),其空间位置与大分子的(M) 根据已知的方法测定本身,一个或多个photoakitvierbare双官能交联剂(C)具有在(1(2共价的范围内的相应的激励频率(交联剂的非光活化端S3(C,C”,C之间 “Lichtausschiuss下大分子(M)的结合),以及适合的官能团(M),所述大分子(M)的频率间隔上述(1个照射的双(2-(具有频率Q,其中(使用strahlungsioser传输福斯特转移)到相邻的交叉 接头(C和/或C“)激活交联剂的可光活化的端部(A和/或A“)(C和/或C”)与大分子(M)的表面发生反应,并与Oberflä 在Abhähgigkeit荧光配体(F)的距离进行反应的分子(M),并通过生物分析方法鉴定的成对连接的基团的表面。