Immunoblot-based activity assay for heme-containing histidine kinases.

ABSTRACT: Histidine kinases (HKs) are essential bacterial signal transduction proteins and attractive drug targets due to their critical cellular functions. Direct measurement of their autophosphorylation activity is crucial for developing inhibitors and advancing disease treatments. While [γ-³²P]-ATP radiolabeling has long been a conventional method for kinase activity measurements, its reliance on ³²P introduces inherent limitations. The isotope's short half-life imposes time-sensitive constraints on experiments, and stringent radiation safety and compliance requirements significantly increase operational costs and hinder scalability. Several alternative methods, including fluorescent phosphate-binding probes and fluorescence- or luminescence-based antibodies, have been devel

SUBMITTER: Larson G 

PROVIDER: S-EPMC12154586 | biostudies-literature | 2025 May

REPOSITORIES: biostudies-literature