兰州大学机构库 >生命科学学院
Strecker合成法合成降冰片烯基甘氨酸及与四嗪的生物正交反应
Alternative TitleStrecker synthesis of 2-amino-2-(bicyclo[2.2.1]hept-5-en-2-yl)acetic acid and bioorthogonal reaction with tetrazine
张紊玮
Thesis Advisor董守良
2010-05-20
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword降冰片烯 四嗪 Strecker合成法 生物正交反应 逆电子需求狄尔斯—阿尔德反应
Abstract目的:降冰片烯与四嗪能够发生逆电子需求狄尔斯—阿尔德反应,放出氮气最终生成二氢哒嗪,符合典型的生物正交反应。与以往的生物正交反应相比,反应具有更高速率、无需任何催化剂、无毒性、底物合成方法简单等特点,但是,还未曾在体内/外标记多肽或蛋白中崭露头角。若将降冰片烯基整合到氨基酸中,进而整合到多肽或蛋白中,再与四嗪进行专一性结合,那么可以实现在生物体内/外对多肽或蛋白的标记。 方法:运用斯特雷克氨基酸合成法 (Strecker synthesis)合成5-降冰片烯基-2-甘氨酸;用肾酰基转移酶 I 分离L-5-降冰片烯基-2-甘氨酸;合成3, 6-二苯甲酸-1, 2, 4, 5-四嗪、3-苄胺-1, 2, 4, 5-四嗪,研究与5-降冰片烯基-2-甘氨酸的环化加成反应;将5-降冰片烯基-2-甘氨酸Fmoc保护,采用Fmoc固相合成法将5-降冰片烯基-2-甘氨酸整合到抹香鲸肌红蛋白一段五肽中,在PBS缓冲液中用四嗪进行标记; 结果与结论:通过Strecker氨基酸合成法,方便合成5-降冰片烯基-2-甘氨酸,总产率41%;用肾酰基转移酶 I分离L-5-降冰片烯基-2-甘氨酸,产率21%;合成3, 6-二苯甲酸-1, 2, 4, 5-四嗪、3-苄胺-1, 2, 4, 5-四嗪,产率分别为67%和23%;用Fmoc固相合成法将5-降冰片烯基-2-甘氨酸整合到抹香鲸肌红蛋白一段五肽中,在PBS缓冲液中用3-苄胺-1, 2, 4, 5四嗪进行了专一性标记。
Other AbstractObjective: Norbornene reacts with tetrazine forming dihydropyridazine products and dinitrogen based on inverse electron demand Diels-Alder reaction. The reaction meets bioorthogonal reaction criteria. Comparing with former bioorthogonal reactions, tetrazine-based cycloadditions has more features such as higher rates, without need of any catalyst, non-toxicity and readily available starting materials, but it still not emerging in labeling peptides or proteins in vivo/in vitro. If norbornene was incorporated into amino acid,and then integrated into peptides or proteins which then were labeled with tetrazine. So we could localize target peptides or protein in vivo/in vitro specifically. Methods: 5-norbornene-2-glycine was synthesized via Strecker synthesis; Kidney acylase I was used to isolate L-5-norbornene-2-glycine from DL-5-norbornene-2-glycine; 3,6-dibenzoic acid-1,2,4,5-tetrazine and 3-benzylamino-1,2,4,5-tetrazine were synthesized separately, then study the cycloaddition reactions between 5-norbornene-2-glycine and tetrazine; integrate 5-norbornene-2-glycine (Fmoc protected) into pentapeptide of sperm whale myoglobin and label pentapeptide using tetrazine in PBS buffer. Results and Conclusion: We synthesized 5-norbornene-2-glycine through Strecker reaction with total yield of 41%, separated L-5-norbornene-2-glycine using Kidney acylase with yield of 21%; Then we synthesized 3,6-dibenzoic acid-1,2,4,5-tetrazine and 3-benzylamino-1,2,4,5-tetrazine with yield of 67% and 23%, respectively; We used Fmoc solid phase peptide synthesis intergrated Fmoc-5-norbornene-2-glycine into pentapeptide of sperm whale myoglobin and labeled pentapeptide using 3-benzylamino-1,2,4,5-tetrazine in PBS buffer.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/221848
Collection生命科学学院
Recommended Citation
GB/T 7714
张紊玮. Strecker合成法合成降冰片烯基甘氨酸及与四嗪的生物正交反应[D]. 兰州. 兰州大学,2010.
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