Contreras-Aguilar María D, Mateo Sandra V, Tecles Fernando, Hirtz Christophe, Escribano Damián, Cerón Jose J
Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, 30100 Murcia, Spain.
IRMB-PPC, CHU Montpellier, Montpellier University, 34295 Montpellier, France.
Biology (Basel). 2021 Mar 16;10(3):227. doi: 10.3390/biology10030227.
This study aimed to evaluate the changes in the activity of total salivary alpha-amylase (TsAA) and both the non-glycosylated and glycosylated salivary alpha-amylase proteoforms (NGsAA and GsAA, respectively) in physical and psychological stress models, estimated using a simple and easily set-up method. The method used was a spectrophotometric assay with 2-chloro-4-nitrophenyl-α-D-maltotriose (CNPG3) as a substrate, incubated with Concanavalin A (ConA) to remove most of the glycosylated protein from the sample. This method allowed the measurement of TsAA and estimation of NGsAA and GsAA activities with imprecision lower than 10%. When this method was applied to two different stress models, differences in the responses of the proteoforms were observed, with the NGsAA activity showing changes of higher magnitude after stress induction than the GsAA activity, and the highest correlation with the State-Trait Anxiety Inventory Scale in the Trier Social Stress Test (TSST). In conclusion, the activity of the two main sAA proteoforms can be easily estimated in saliva, and their measurement can provide additional information on TsAA activity in physical or psychological stress situations.
本研究旨在评估在物理和心理应激模型中，使用一种简单且易于设置的方法估计的总唾液α-淀粉酶（TsAA）以及非糖基化和糖基化唾液α-淀粉酶蛋白亚型（分别为NGsAA和GsAA）的活性变化。所采用的方法是一种分光光度测定法，以2-氯-4-硝基苯基-α-D-麦芽三糖（CNPG3）为底物，并与伴刀豆球蛋白A（ConA）一起孵育，以从样品中去除大部分糖基化蛋白。该方法能够测量TsAA，并估计NGsAA和GsAA的活性，不精密度低于10%。当将此方法应用于两种不同的应激模型时，观察到蛋白亚型反应的差异，应激诱导后NGsAA活性的变化幅度高于GsAA活性，且在特里尔社会应激测试（TSST）中与状态-特质焦虑量表的相关性最高。总之，两种主要唾液α-淀粉酶蛋白亚型的活性可以在唾液中轻松估计，它们的测量可为物理或心理应激情况下的TsAA活性提供额外信息。
