Department of Medical Biochemistry, University of Gothenburg, Gothenburg, Sweden.
Department of Medical Biochemistry, University of Gothenburg, Gothenburg, Sweden.
Anal Biochem. 2020 May 15;597:113668. doi: 10.1016/j.ab.2020.113668. Epub 2020 Mar 25.
In order to demonstrate transglutaminase activity in biological samples immunological as well as glutamine- and amine-donor based assays are commonly used. However, the identification of the transglutaminase reaction product, i. e. the isopeptide cross-linked peptides/proteins or the deamidated protein/peptide are often neglected. This article describes a workflow for the detection of the products of transglutaminase-catalyzed reactions. In particular, possible pitfalls and traps that can arise during the mass spectrometry-based identification of isopeptide cross-links are addressed and characterised on actual samples.
为了在生物样本中展示转谷氨酰胺酶的活性，免疫测定法以及基于谷氨酰胺和胺供体的测定法通常被使用。然而，转谷氨酰胺酶反应产物（即，异肽交联肽/蛋白质或脱酰胺蛋白质/肽）的鉴定经常被忽视。本文描述了一种检测转谷氨酰胺酶催化反应产物的工作流程。特别地，在基于质谱的异肽交联鉴定过程中可能出现的潜在问题和陷阱，以及在实际样本中的特征都被进行了探讨。
