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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.

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Identifying Inhibitor Targets in Mycobacteria by Activity-Based Probe Profiling

  • 实验方案

  • 发表日期：

    2025年

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  • Neetika Jaisinghani,
  • Isabel Sakarin,
  • Hiren V. Patel,
  • Michael Li,
  • Mary L. Previti,
  • Jessica C. Seeliger

摘要

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.