Site-specific protein transamination using N-methylpyridinium-4-carboxaldehyde.
Site-specific protein transamination using N-methylpyridinium-4-carboxaldehyde.
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DOI：
10.1021/ja408868a
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发表时间：
2013-11-13
影响因子：
15
通讯作者：
Francis MB
中科院分区：
文献类型：
--
作者：
Witus LS;Netirojjanakul C;Palla KS;Muehl EM;Weng CH;Iavarone AT;Francis MB
合成基团与蛋白质的受控连接对于许多领域都很重要，包括治疗学，其中抗体-药物缀合物是生物药物的新兴领域。我们以前已经报道了一种位点特异性的蛋白质修饰方法，使用转氨反应，化学选择性氧化的N-末端胺的多肽链的酮或醛基。新引入的羰基可用于通过形成肟或腙键在一个位置与合成基团缀合。为了扩大该反应的范围，我们使用了组合肽库筛选平台作为探索新的转氨试剂的方法，同时确定其最佳N-末端序列。N-甲基吡啶鎓-4-甲醛苯磺酸盐（Rapoport盐，RS）被鉴定为当与谷氨酸末端肽和蛋白质配对时的高效转氨试剂。这一发现确定RS是一种特别适合抗体修饰的转氨试剂。使用已知的治疗性抗体赫赛汀，证明RS可用于修饰野生型抗体的重链，或在N-末端序列突变后修饰重链和轻链以添加谷氨酸残基。
The controlled attachment of synthetic groups to proteins is important for a number of fields, including therapeutics, where antibody-drug conjugates are an emerging area of biologic medicines. We have previously reported a site-specific protein modification method using a transamination reaction that chemoselectively oxidizes the N-terminal amine of a polypeptide chain to a ketone or an aldehyde group. The newly introduced carbonyl can be used for conjugation to a synthetic group in one location through the formation of an oxime or a hydrazone linkage. To expand the scope of this reaction, we have used a combinatorial peptide library screening platform as a method to explore new transamination reagents while simultaneously identifying their optimal N-terminal sequences. N-methylpyridinium-4-carboxaldehyde benzenesulfonate salt (Rapoport's salt, RS) was identified as a highly effective transamination reagent when paired with glutamate-terminal peptides and proteins. This finding establishes RS as a transamination reagent that is particularly well suited for antibody modification. Using a known therapeutic antibody, herceptin, it was demonstrated that RS can be used to modify the heavy chains of the wild type antibody, or both the heavy and the light chains after N-terminal sequence mutation to add glutamate residues.
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影响因子：
15
作者：
Scheck, Rebecca A.;Dedeo, Michel T.;Francis, Matthew B.
通讯作者：
Francis, Matthew B.
影响因子：
5.6
作者：
Harris, LJ;Skaletsky, E;McPherson, A
通讯作者：
McPherson, A
影响因子：
50.3
作者：
Neve, Richard M.;Chin, Koei;Gray, Joe W.
通讯作者：
Gray, Joe W.
影响因子：
4.7
作者：
Baker, DP;Lin, EY;Pepinsky, RB
通讯作者：
Pepinsky, RB
影响因子：
64.8
作者：
Cho, HS;Mason, K;Leahy, DJ
通讯作者：
Leahy, DJ