Investigating Glyoxylate-Mediated Transamination Using Dipeptide Arrays and Proteomic Peptide Mixtures,Bioconjugate Chemistry

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Investigating Glyoxylate-Mediated Transamination Using Dipeptide Arrays and Proteomic Peptide Mixtures
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2018-09-20 00:00:00 , DOI: 10.1021/acs.bioconjchem.8b00475
Xiaohong Tan ₁ , Chuan-Fa Liu ₂

Affiliation

Glyoxylate-mediated transamination (GT) is a classic, potentially general, and N-terminus-specific protein modification method useful for the preparation of bioconjugates. However, there is a lack of information on whether and how readily a particular N-terminal amino acid (in the context of a peptide chain) can be converted to the 2-oxoacyl moiety under GT conditions. Here, we conducted a systematic investigation of GT using membrane-bound dipeptide arrays that include all the 400 possible dipeptide combinations of the 20 genetically encoded amino acids. This colorimetric method offers a convenient way to assess the GT reaction tendency of N-terminal residues by the naked eye. It also provides interesting information about the effect of the second residues on GT, which has not been reported previously. In addition, we also designed a proteomics approach to study GT in solution using tryptic peptide mixtures, which not only confirmed many of our findings in peptide array assays but also revealed potential side reaction products. Taken together, our studies will make the future use of GT for protein modification in a much more predictable way.

中文翻译：

研究使用二肽阵列和蛋白质组肽混合物的乙醛酸介导的转氨作用

乙醛酸酯介导的转氨作用（GT）是一种经典的，可能通用的N端特异的蛋白质修饰方法，可用于制备生物缀合物。然而，缺乏关于在GT条件下是否以及如何容易地将特定的N-末端氨基酸（在肽链的情况下）转化为2-氧代酰基部分的信息。在这里，我们使用膜结合的二肽阵列进行了GT的系统研究，该阵列包括20种遗传编码氨基酸的所有400种可能的二肽组合。此比色法提供了一种方便的方法，可以通过肉眼评估N末端残基的GT反应趋势。它还提供了有关第二个残基对GT的影响的有趣信息，以前尚未报道过。此外，我们还设计了一种蛋白质组学方法来研究使用胰蛋白酶肽混合物的溶液中的GT，这不仅证实了我们在肽阵列测定中的许多发现，而且还揭示了潜在的副产物。综上所述，我们的研究将以更可预测的方式使GT将来用于蛋白质修饰。

更新日期：2018-09-20

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