Xi Ying, Collins Leonard B, Bai Hongxia, Biehl Andreea, Mora-Navarro Camilo, Freytes Donald, Islam Williams Taufika
Department of Chemistry, North Carolina State University, Raleigh, NC, USA.
Molecular Education, Technology and Research Innovation Center (METRIC), North Carolina State University, Raleigh, NC, USA.
Methods Mol Biol. 2025;2884:179-191. doi: 10.1007/978-1-0716-4298-6_12.
Extracellular matrix (ECM) from decellularized mammalian tissues has been used in many therapeutic applications. The tissue-specific composition of the ECM is critically associated with therapeutic performance. However, ECM translation needs to be improved because of the complex composition and limited understanding of ECM repairing mechanisms due partly to incomplete proteomic interrogation of ECM samples. In this chapter, we describe a multi-enzyme, bottom-up proteomics workflow employing trypsin, Lys-C, collagenase, and elastase to enhance the digestion of ECM and increase total protein coverage. The outcomes from the reported approach, in a standardized manner, enable users to pinpoint changes in the ECM composition, thereby facilitating the establishment of mechanistic correlations between ECM composition and its effects.
来自脱细胞哺乳动物组织的细胞外基质（ECM）已被用于许多治疗应用。ECM的组织特异性组成与治疗性能密切相关。然而，由于ECM组成复杂且对其修复机制的理解有限（部分原因是对ECM样本的蛋白质组学分析不完整），ECM的转化应用仍需改进。在本章中，我们描述了一种多酶自下而上的蛋白质组学工作流程，该流程使用胰蛋白酶、Lys-C、胶原酶和弹性蛋白酶来增强ECM的消化并增加总蛋白覆盖率。所报道方法的结果以标准化方式使用户能够确定ECM组成的变化，从而有助于建立ECM组成与其效应之间的机制关联。
