用于通过MRM-MS定量肌球蛋白磷酸化的工具和方案。
Tools and protocol for quantification of myosin phosphorylation with MRM-MS.
作者信息
MacDonald Justin A, Ulke-Lemée Annegret, Chappellaz Mona, Segboer Hayden
机构信息
Department of Biochemistry & Molecular Biology, Cumming School of Medicine, University of Calgary, Calgary, AB, T2N 4Z6, Canada.
出版信息
MethodsX. 2018 May 17;5:466-474. doi: 10.1016/j.mex.2018.05.004. eCollection 2018.
The phosphorylation of myosin regulatory light chain (LC20) at Thr18 and Ser19 is positively correlated with tension development in smooth muscle tissue, and the molar stoichiometry of LC20 phosphorylation is commonly profiled as a measure of smooth muscle contractility. We provide details for a newly applied multiple reaction monitoring (MRM)-mass spectrometry (MS) method for the quantification of LC20 phosphorylation at Thr18 and Ser19. This MRM-MS method provides a robust alternative to antibody-based detection systems (such as Phos-Tag SDS-PAGE) for the quantification of LC20 phosphorylation.
肌球蛋白调节轻链（LC20）在苏氨酸18和丝氨酸19位点的磷酸化与平滑肌组织中的张力发展呈正相关，并且LC20磷酸化的摩尔化学计量通常被用作衡量平滑肌收缩力的指标。我们详细介绍了一种新应用的多反应监测（MRM）-质谱（MS）方法，用于定量苏氨酸18和丝氨酸19位点的LC20磷酸化。这种MRM-MS方法为基于抗体的检测系统（如Phos-Tag SDS-PAGE）在定量LC20磷酸化方面提供了一种可靠的替代方法。
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