Clinic Analysis Interdisciplinary Laboratory (Interlab-UMU), Campus of Excellence Mare Nostrum, Murcia, Spain.
LBPC/PPC - IRMB, CHU de Montpellier, INSERM, Montpellier University, 80 rue Augustin Fliche, Montpellier, France.
Clin Oral Investig. 2020 Jan;24(1):475-486. doi: 10.1007/s00784-019-03021-9. Epub 2019 Aug 6.
To evaluate the sAA proteoforms' expression during different stimulation situations.
This study evaluated the salivary alpha-amylase (sAA) proteoforms' behavior by western blot (WB) analysis and high-resolution mass spectrometry (LC-MS/MS) in different situations that produce increases in sAA activity. For this purpose, six healthy women with a similar body mass index, age, and fit, underwent different sAA stimulation tests, such as acetic acid stimulation, psychological stress using the standardized Trier social stress test, and physical effort using the Cooper treadmill test.
The three models showed an increase in sAA activity. The WB demonstrated seven common bands observed in the six women (band one at 59 kDa, two at 56 kDa, three at 48 kDa, four at 45 kDa, five at 41 kDa, six at 36 kDa, and seven at 14 kDa), in which sAA protein was identified. The individual WB analysis showed that band two, which corresponded to the native non-glycosylated sAA proteoform, had a higher increase after the three sAA stimulation inducers, and this band was also the only proteoform correlated with sAA activity (r = 0.56, P = 0.001). In addition, when the label-free quantification analysis was performed, the different proteoforms showed different responses depending on the type of stimulation.
This preliminary study showed that the diverse sAA proteoforms' expression depends on the different stimulation models.
This study opens new perspectives and challenges for the use of the different alpha-amylase proteoforms as possible biomarkers in addition to the sAA activity.
评估不同刺激情况下唾液淀粉酶（sAA）蛋白水解物的表达。
本研究通过western blot（WB）分析和高分辨率质谱（LC-MS/MS）评估了不同情况下 sAA 活性增加时唾液α-淀粉酶（sAA）蛋白水解物的行为。为此，6 名身体质量指数、年龄和健康状况相似的健康女性接受了不同的 sAA 刺激测试，如醋酸刺激、使用标准化的特里尔社会应激测试的心理压力和使用库珀跑步机测试的体力消耗。
三种模型均显示 sAA 活性增加。WB 显示了在六名女性中观察到的七种常见条带（一条位于 59 kDa，两条位于 56 kDa，三条位于 48 kDa，四条位于 45 kDa，五条位于 41 kDa，六条位于 36 kDa，七条位于 14 kDa），其中鉴定出 sAA 蛋白。个体 WB 分析表明，对应于天然非糖基化 sAA 蛋白水解物的第二条带在三种 sAA 刺激诱导物后增加幅度更高，并且该条带也是与 sAA 活性唯一相关的蛋白水解物（r=0.56，P=0.001）。此外，当进行无标记定量分析时，不同的蛋白水解物表现出不同的反应，这取决于刺激的类型。
这项初步研究表明，不同的 sAA 蛋白水解物的表达取决于不同的刺激模型。
这项研究为使用不同的α-淀粉酶蛋白水解物作为 sAA 活性的可能生物标志物开辟了新的视角和挑战。
