Jaisinghani Neetika, Sakarin Isabel, Patel Hiren V, Li Michael, Previti Mary L, Seeliger Jessica C
Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY, USA.
Department of Microbiology and Immunology, Stony Brook University, Stony Brook, NY, USA.
Methods Mol Biol. 2025;2921:139-154. doi: 10.1007/978-1-0716-4502-4_7.
Activity-based protein profiling (ABPP) enables the detection of protein reactivity across entire proteomes. Here, we describe the application of ABPP to the identification of serine hydrolase inhibitor targets in Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis in humans. We highlight the adaptation of stable isotope labeling of amino acids (SILAC) to Mtb using a modified growth medium with an isotopically labeled sole nitrogen source and detail the resulting special requirements for proteomics analysis.
基于活性的蛋白质谱分析（ABPP）能够检测整个蛋白质组中的蛋白质反应性。在此，我们描述了ABPP在鉴定结核分枝杆菌（Mtb，人类结核病的病原体）中丝氨酸水解酶抑制剂靶点方面的应用。我们着重介绍了利用含有同位素标记单一氮源的改良生长培养基，将氨基酸稳定同位素标记（SILAC）应用于结核分枝杆菌，并详细说明了由此产生的蛋白质组学分析的特殊要求。
