Visual Analysis and Inhibitor Screening of Leucine Aminopeptidase, a Key Virulence Factor for Pathogenic Bacteria-Associated Infection

Leucine aminopeptidase (LAP) is a hydrolase for the hydrolysis of peptides or proteins containing a leucine residue at the N-terminal. It is also known to be a key virulence factor for the pathogenic abilities of various pathogens causing infectious diseases, which indicated a new insight into the diagnosis and therapy of pathogenic infections. A new fluorescent probe (S)-2-amino-N-(4-(((6,8-dichloro-9,9-dimethyl-7-oxo-7,9-dihydroacridin-2-yl)oxy)methyl)phenyl)-4-methylpentanamide (DDBL) containing DDAO as the fluorophore and leucine as the recognition group was developed for LAP. By real-time visual sensing of LAP, six bacteria with LAP expression were identified efficiently from human feces, as well as by sensitive visual analysis using native-PAGE specially stained with DDBL. Furthermore, a high throughput screening system established with DDBL was applied to identify a natural inhibitor (3-acetyl-11-keto-beta-boswellic acid, AKBA), which could attenuate mouse sepsis induced by Staphylococcus aureus. Therefore, the visual sensing of LAP by DDBL suggested the application for target bacteria identification and LAP homolog analysis as well as potential inhibitor expounding for treatment of bacterial infections.

Automated summary

Leucine aminopeptidase (LAP) is a key hydrolytic enzyme for specific peptides and proteins, and is known as a crucial virulence factor for pathogenic infections. The development of a new fluorescent probe DDBL allows real-time detection of LAP expression in bacteria from feces, and is also used for high-throughput screening to identify inhibitors.