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    • 4. 发明授权
    • Labelling of fusion proteins with synthetic probes
    • 用合成探针标记融合蛋白
    • US08227602B2
    • 2012-07-24
    • US12309554
    • 2007-07-24
    • Arnaud GautierKai JohnssonMaik KindermannAlexandre JuilleratFlorent Beaufils
    • Arnaud GautierKai JohnssonMaik KindermannAlexandre JuilleratFlorent Beaufils
    • C07D239/02
    • G01N33/581C07D239/47C07D403/14C07D405/12C07D487/04C12N9/12C12Q1/48
    • The invention relates to new proteins called alkylcytosine transferases (ACTs) derived from O6-alkylguanine-DNA alkyltransferase, and to substrates for ACTs specifically transferring a label to these ACTs and to fusion proteins comprising these. The substrates according of the invention are substituted cytosines of formula (I) wherein R1 is an aromatic or a heteroaromatic group, or an optionally substituted unsaturated alkyl, cycloalkyl or heterocyclyl group with the double bond connected to OCH2—; R2 is a linker; and L is a label or a plurality of same or different labels. The invention further relates to methods of transferring label L from these substrates of formula (I) to ACTs and ACT fusion proteins. The system of ACT-compound of formula (I) is particularly suitable for double labelling studies together with the known system O6-alkylguanine-DNA alkyltransferase (AGT)-benzylguanines.
    • 本发明涉及从O6-烷基鸟嘌呤-DNA烷基转移酶衍生的称为烷基胞嘧啶转移酶(ACT)的新蛋白质,以及特异性将标记转移至这些ACT的ACT的底物和包含它们的融合蛋白。 根据本发明的底物是其中R 1是芳族或杂芳族基团的式(I)的取代的胞嘧啶或具有与OCH 2连接的双键的任选取代的不饱和烷基,环烷基或杂环基; R2是连接体; L是标签或多个相同或不同的标签。 本发明还涉及将标记L从这些式(I)底物转移到ACT和ACT融合蛋白的方法。 式(I)的ACT-化合物的系统特别适用于已知的系统O6-烷基鸟嘌呤-DNA烷基转移酶(AGT) - 苄基胍的双重标记研究。
    • 5. 发明授权
    • Specific substrates for O6-alkylguanine-DNA alkyltransferase
    • O6-烷基鸟嘌呤-DNA烷基转移酶的特异性底物
    • US08163479B2
    • 2012-04-24
    • US10591162
    • 2005-03-01
    • Hughes JaccardKai JohnssonMaik KindermannIndia Christina Sielaff
    • Hughes JaccardKai JohnssonMaik KindermannIndia Christina Sielaff
    • C07D473/18C07D519/00C07D495/04C07D473/40
    • C07D473/18
    • The invention relates to substrates for O6-alkylguanine-DNA alkyltransferases (AGT) of formula R1-A-X—CH2—R3—R4-L1, wherein A is a group recognized by AGT as a substrate, X is oxygen or sulfur, R1 is a group —R2-L2 or a group R5, R2 and R4 are, independently of each other, a linker, R3 is an aromatic or a heteroaromatic group, or an optionally substituted unsaturated alkyl, cycloalkyl or heterocyclyl group with the double bond connected to CH2, R5 is arylmethyl or heteroarylmethyl or an optionally substituted cycloalkyl, cycloalkenyl or heterocyclyl group, L1 is a label, a plurality of same or different labels, a bond connecting R4 to A forming a cyclic substrate, or a further group —R3—CH2—X-A-R1, and L2 is a label or a plurality of same or different labels. The invention further relates to methods of transferring a label from these substrates to O6-alkylguanine-DNA alkyltransferases (AGT) and AGT fusion proteins.
    • 本发明涉及式R1-AX-CH2-R3-R4-L1的O6-烷基鸟嘌呤-DNA烷基转移酶(AGT)的底物,其中A为被AGT识别为底物的基团,X为氧或硫,R1为 基团-R2-L2或基团R5,R2和R4彼此独立地是连接基,R3是芳族或杂芳族基团,或任选取代的不饱和烷基,环烷基或杂环基,其双键与CH2连接 R 5是芳基甲基或杂芳基甲基或任选取代的环烷基,环烯基或杂环基,L1是标记,多个相同或不同的标记,连接R4与A形成环状底物的键或另外的基团-R 3 -CH 2 - XA-R1和L2是标签或多个相同或不同的标签。 本发明还涉及将标记从这些底物转移到O6-烷基鸟嘌呤-DNA烷基转移酶(AGT)和AGT融合蛋白的方法。