在线毛细管电泳-质谱分析组蛋白-5及其降解产物。
Online capillary electrophoresis - mass spectrometry analysis of histatin-5 and its degradation products.
机构信息
CMP Scientific, Corp., 760 Parkside Ave, STE 211, Brooklyn, NY 11226, USA.
Department of Chemical and Biomolecular Engineering, University of Maryland, College Park, MD, USA.
出版信息
Analyst. 2020 Jul 13;145(14):4787-4794. doi: 10.1039/d0an00756k.
Histatin-5 (Hst-5) is a human salivary peptide with antibacterial and antifungal activities. Thorough characterization and reliable quantification of Hst-5 and its degradation products are essential for understanding the Hst-5 degradation pathway. Due to the highly basic and strong cationic nature of the Hst-5 peptide, the quantitative analysis of Hst-5 and its degradation forms by online mass spectrometry remains challenging. Here, we adopt a recently developed electrokinetically pumped sheath liquid capillary electrophoresis - mass spectrometry (CE-MS) coupling technology, and successfully apply it for the analysis of Hst-5 and its degradation products. Our CE-MS method is demonstrated to be robust and quantitative. This novel analytical platform is reproducible and free of sample carryover. The efficacy of this method is demonstrated with a kinetic study of Hst-5 degradation by Sap9, a secreted aspartic peptidase. Our work demonstrates the potential of online CE-MS as a powerful approach for characterizing highly basic peptides.
Histatin-5（Hst-5）是一种具有抗菌和抗真菌活性的人类唾液肽。彻底表征和可靠定量 Hst-5 及其降解产物对于了解 Hst-5 降解途径至关重要。由于 Hst-5 肽具有高度碱性和强阳离子性质，因此通过在线质谱法对 Hst-5 及其降解形式进行定量分析仍然具有挑战性。在这里，我们采用了最近开发的电动泵送鞘液毛细管电泳 - 质谱（CE-MS）联用技术，并成功地将其应用于 Hst-5 及其降解产物的分析。我们的 CE-MS 方法被证明是稳健和定量的。这种新的分析平台具有重现性且无样品残留。通过 Sap9（一种分泌的天冬氨酸肽酶）对 Hst-5 降解的动力学研究证明了该方法的有效性。我们的工作证明了在线 CE-MS 作为一种强大的方法来表征高度碱性肽的潜力。
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